1. B Y T S: 26 /Q-BYT CNG HA X HI CH NGHA VIT NAM c lp - T do - Hnh phc H Ni, ngy 03 thng 01 nm 2013 QUYT NH V vic ban hnh ti liu Hng dn quy trnh k thut chuyn ngnh Vi sinh Y hc B TRNG B Y T Cn c Lut khm bnh, cha bnh nm 2009; Cn c Ngh nh s 63/2012/N-CP ngy 31/8/2012 ca Chnh Ph quy nh chc nng, nhim v, quyn hn v c cu t chc ca B Y t; Xt Bin bn hp ca Hi ng nghim thu Hng dn Quy trnh k thut khm bnh, cha bnh chuyn ngnh Vi sinh Y hc ca B Y t; Theo ngh ca Cc trng Cc Qun l Khm, cha bnh, QUYT NH: iu 1. Ban hnh km theo Quyt nh ny ti liu Hng dn quy trnh k thut chuyn ngnh Vi sinh Y hc, gm 231 quy trnh k thut. iu 2. Ti liu Hng dn quy trnh k thut chuyn ngnh Vi sinh Y hc ban hnh km theo Quyt nh ny c p dng ti cc c s khm bnh, cha bnh. Cn c vo ti liu hng dn ny v iu kin c th ca n v, Gim c c s khm bnh, cha bnh xy dng v ban hnh ti liu Hng dn Quy trnh k thut Vi sinh Y hc ph hp thc hin ti n v. iu 3. Quyt nh ny c hiu lc k t ngy k ban hnh. iu 4. Cc ng, b: Chnh Vn phng B, Chnh Thanh tra B, Cc trng Cc Qun l Khm, cha bnh, Cc trng v V trng cc Cc, V thuc B Y t, Gim c cc bnh vin, vin c ging bnh trc thuc B Y t, Gim c S Y t cc tnh, thnh ph trc thuc trung ng, Th trng Y t cc B, Ngnh v Th trng cc n v c lin quan chu trch nhim thi hnh Quyt nh ny./. Ni nhn: - Nh iu 4; - B trng B Y t ( b/c); - Cc Th trng BYT; - Bo him X hi Vit Nam ( phi hp); - Cng thng tin in t BYT; - Website Cc KCB; - Lu VT, KCB. KT. B TRNG TH TRNG k Nguyn Th Xuyn 2. B Y T CNG HA X HI CH NGHA VIT NAM c lp - T do- Hnh Phc DANH SCH CC QUY TRNH K THUT CHUYN NGNH VI SINH HC (Ban hnh km theo Quyt nh s: 26/Q-BYT ngy 03 thng 01 nm 2014 ca B trng B Y t) TT TN QUY TRNH K THUT Danh mc xt nghim vi khun A. Vi khun chung 1 Vi khun nhum soi 2 Vi khun test nhanh 3 Vi khun nui cy v nh danh phng php thng thng 4 Vi khun nui cy v nh danh h thng t ng 5 Vi khun nui cy, nh danh v khng thuc h thng t ng 6 Vi khun khng thuc nh tnh 7 Vi khun khng thuc h thng t ng 8 Vi khun khng thuc nh lng (MIC) (cho 1 loi khng sinh) 9 Vi khun khng sinh phi hp 10 Vi khun k kh nui cy v nh danh 11 Vi khun khng nh 12 Vi khun nh danh PCR 13 Vi khun nh danh gii trnh t gene 14 Vi khun khng thuc PCR 15 Vi khun khng thuc gii trnh t gene 16 Vi h ng rut B. Mycobacteria 17 AFB trc tip nhum Ziehl-Neelsen 18 AFB trc tip nhum hunh quang 19 Mycobacterium tuberculosis nui cy mi trng lng 20 Mycobacterium tuberculosis nui cy mi trng c 21 Mycobacterium tuberculosis Mantoux 22 Mycobacterium tuberculosis khng thuc hng 1 mi trng c 23 Mycobacterium tuberculosis khng thuc hng 1 mi trng lng 24 Mycobacterium tuberculosis khng thuc hng 2 mi trng c 25 Mycobacterium tuberculosis khng thuc hng 2 mi trng lng 26 Mycobacterium tuberculosis khng thuc PZA mi trng lng 27 Mycobacterium tuberculosis pyrazinamidase 28 Mycobacterium tuberculosis nh danh v khng RMP Xpert 29 Mycobacterium tuberculosis a khng LPA 30 Mycobacterium tuberculosis siu khng LPA 31 Mycobacterium tuberculosis PCR h thng t ng 32 Mycobacterium tuberculosis Real-time PCR 33 Mycobacterium tuberculosis spoligotyping 3. 34 Mycobacterium tuberculosis RFLP typing 35 NTM (Non tuberculosis mycobacteria) nui cy mi trng lng 36 NTM (Non tuberculosis mycobacteria) nui cy mi trng c 37 NTM (Non tuberculosis mycobacteria) nh danh LPA 38 Mycobacterium leprae nhum soi 39 Mycobacterium leprae PCR 40 Mycobacterium leprae mnh sinh thit C. Vibrio cholerae 41 Vibrio cholerae nhum soi 42 Vibrio cholerae nhum hunh quang 43 Vibrio cholerae nui cy, nh danh v khng thuc 44 Vibrio cholerae PCR 45 Vibrio cholerae gii trnh t gene D. Neisseria gonorrhoeae 46 Neisseria gonorrhoeae nhum soi 47 Neisseria gonorrhoeae nui cy, nh danh v khng thuc 48 Neisseria gonorrhoeae PCR 49 Neisseria gonorrhoeae Real-time PCR E. Neisseria meningitidis 50 Neisseria meningitidis nhum soi 51 Neisseria meningitidis nui cy, nh danh v khng thuc 52 Neisseria meningitidis PCR 53 Neisseria meningitidis Real-time PCR F. Cc vi khun khc 54 Chlamydia test nhanh 55 Chlamydia nhum hunh quang 56 Chlamydia Ab min dch bn t ng 57 Chlamydia PCR 58 Chlamydia Real-time PCR 59 Chlamydia Real-time PCR h thng t ng 60 Clostridium nui cy, nh danh 61 Clostridium difficile min dch bn t ng 62 Clostridium difficile PCR 63 Leptospira test nhanh 64 Mycoplasma pneumoniae Real-time PCR 65 Mycoplasma hominis nui cy, nh danh v khng thuc 66 Rickettsia Ab min dch bn t ng 67 Salmonella Widal 68 Streptococcus pyogenes ASO 69 Treponema pallidum soi ti 70 Treponema pallidum nhum soi 71 Treponema pallidum RPR nh tnh v nh lng 72 Treponema pallidum TPHA nh tnh v nh lng 73 Ureaplasma urealyticum nui cy, nh danh v khng thuc Danh mc xt nghim virus 4. A. Virus chung 74 Virus test nhanh 75 Virus Ag min dch bn t ng 76 Virus Ab min dch bn t ng B. Hepatitis virus 77 HBsAg test nhanh 78 HBsAg min dch bn t ng 79 HBsAg min dch t ng 80 HBsAg khng nh 81 HBsAg nh lng 82 HBsAb min dch bn t ng 83 HBsAb nh lng 84 HBc IgM min dch bn t ng 85 HBc IgM min dch t ng 86 HBc total min dch bn t ng 87 HBc total min dch t ng 88 HBeAg min dch bn t ng 89 HBeAg min dch t ng 90 HBeAb min dch bn t ng 91 HBeAb min dch t ng 92 HBV o ti lng Real-time PCR 93 HBV o ti lng h thng t ng 94 HBV genotype PCR 95 HBV genotype Real-time PCR 96 HBV genotype gii trnh t gene 97 HBV khng thuc Real-time PCR (cho 1 loi thuc) 98 HBV khng thuc gii trnh t gene 99 HCV Ab min dch bn t ng 100 HCV Ab min dch t ng 101 HCV Ag/Ab min dch bn t ng 102 HCV Core Ag min dch t ng 103 HCV PCR 104 HCV o ti lng Real-time PCR 105 HCV o ti lng h thng t ng 106 HCV genotype Real-time PCR 107 HCV genotype gii trnh t gene 108 HAV IgM min dch bn t ng 109 HAV IgM min dch t ng 110 HAV total min dch bn t ng 111 HAV total min dch t ng 112 HDV Ag min dch bn t ng 113 HDV IgM min dch bn t ng 114 HDV Ab min dch bn t ng 115 HEV IgM test nhanh 116 HEV IgM min dch bn t ng 5. C. HIV 117 HIV Ab test nhanh 118 HIV Ag/Ab test nhanh 119 HIV Ab min dch bn t ng 120 HIV Ag/Ab min dch bn t ng 121 HIV Ag/Ab min dch t ng 122 HIV khng nh 123 HIV o ti lng Real-time PCR 124 HIV o ti lng h thng t ng 125 HIV genotype gii trnh t gene 126 HIV khng thuc gii trnh t gene D. Dengue virus 127 Dengue virus NS1Ag test nhanh 128 Dengue virus NS1Ag/IgM/IgG test nhanh 129 Dengue virus IgM/IgG test nhanh 130 Dengue virus IgM min dch bn t ng 131 Dengue virus IgG min dch bn t ng 132 Dengue virus PCR 133 Dengue virus serotype PCR E. Herpesviridae 134 CMV IgM min dch bn t ng 135 CMV IgM min dch t ng 136 CMV IgG min dch bn t ng 137 CMV IgG min dch t ng 138 CMV PCR 139 CMV Real-time PCR 140 CMV o ti lng h thng t ng 141 CMV Avidity 142 HSV 1+2 IgM min dch bn t ng 143 HSV 1+2 IgG min dch bn t ng 144 HSV Real-time PCR 145 VZV Real-time PCR 146 EBV-VCA IgM min dch bn t ng 147 EBV-VCA IgG min dch bn t ng 148 EBV EA-D IgG min dch bn t ng 149 EBV EB-NA IgG min dch bn t ng 150 EBV PCR 151 EBV Real-time PCR E. Enterovirus 152 EV71 PCR 153 EV71 Real-time PCR 154 EV71 genotype gii trnh t gene 155 Enterovirus PCR 156 Enterovirus genotype gii trnh t gene F. Cc virus khc 6. 157 Adenovirus Real-time PCR 158 BK/JC virus PCR 159 HPV PCR 160 HPV Real-time PCR 161 HPV genotype Real-time PCR 162 HPV genotype PCR h thng t ng 163 HPV genotype gii trnh t gene 164 Influenza virus A, B test nhanh 165 Influenza virus A, B Real-time PCR 166 Influenza virus A, B gii trnh t gene 167 JEV IgM min dch bn t ng 168 Measles virus Ab min dch bn t ng 169 Rotavirus test nhanh 170 RSV Ab min dch bn t ng 171 RSV Real-time PCR 172 Rubella virus IgM min dch bn t ng 173 Rubella virus IgM min dch t ng 174 Rubella virus IgG min dch bn t ng 175 Rubella virus IgG min dch t ng 176 Rubella virus PCR 177 Rubella virus gii trnh t gene Danh mc xt nghim KST A. K sinh trng trong phn 178 Hng cu, bch cu trong phn soi ti 179 Hng cu trong phn test nhanh 180 n bo ng rut soi ti 181 n bo ng rut nhum soi 182 Trng giun, sn soi ti 183 Trng giun soi tp trung 184 Strongyloides stercoralis (giun ln) u trng soi ti B. K sinh trng trong mu 185 Angiostrogylus cantonensis (Giun trn chut) Ab min dch bn t ng 186 Clonorchis/Opisthorchis (Sn l gan nh) Ab min dch bn t ng 187 Cysticercus cellulosae (Sn ln) Ab min dch bn t ng 188 Entamoeba histolytica (Amip) Ab min dch bn t ng 189 Fasciola (Sn l gan ln) Ab min dch bn t ng 190 Filaria (Giun ch) u trng trong mu nhum soi 191 Gnathostoma Ab min dch bn t ng 192 Plasmodium (K sinh trng st rt) nhum soi nh tnh 193 Plasmodium (K sinh trng st rt) nhum soi nh lng 194 Plasmodium (K sinh trng st rt) Ag test nhanh 195 Strongyloides stercoralis (Giun ln) Ab min dch bn t ng 7. 196 Toxocara (Giun a ch, mo) Ab min dch bn t ng 197 Toxoplasma IgM min dch t ng 198 Toxoplasma IgG min dch t ng 199 Toxoplasma Avidity C. K sinh trng ngoi da 200 Demodex soi ti 201 Demodex nhum soi 202 Phthirus pubis (Rn mu) soi ti 203 Phthirus pubis (Rn mu) nhum soi 204 Sarcoptes scabies hominis(Gh) soi ti 205 Sarcoptes scabies hominis(Gh) nhum soi D. K sinh trng trong cc bnh phm khc 206 Cysticercus cellulosae (Sn ln) u trng soi mnh sinh thit 207 Gnathostoma u trng soi mnh sinh thit 208 Pneumocystis jirovecii nhum soi 209 Taenia (Sn dy) soi ti nh danh 210 Toxocara (Giun a ch, mo) soi mnh sinh thit 211 Trichinella spiralis (Giun xon) soi mnh sinh thit 212 Trichomonas vaginalis soi ti 213 Trichomonas vaginalis nhum soi Danh mc xt nghim Vi nm 214 Vi nm soi ti 215 Vi nm test nhanh 216 Vi nm nhum soi 217 Vi nm nui cy v nh danh phng php thng thng 218 Vi nm nui cy v nh danh h thng t ng 219 Vi nm nui cy, nh danh v khng thuc h thng t ng 220 Vi nm khng nh 221 Vi nm khng thuc nh lng (MIC) (cho 1 loi khng sinh) 222 Vi nm PCR 223 Vi nm gii trnh t gene Danh mc xt nghim nh gi nhim khun bnh vin 224 Vi sinh vt cy kim tra khng kh 225 Vi sinh vt cy kim tra bn tay 226 Vi sinh vt cy kim tra dng c tit trng 227 Vi sinh vt cy kim tra b mt 228 Vi sinh vt cy kim tra nc sinh hot 229 Vi sinh vt cy kim tra nc thi 230 Vi khun khng thuc - Pht hin ngi mang 231 Vi khun gy nhim trng bnh vin - Pht hin ngun nhim (Tng s 231 quy trnh k thut) KT. B TRNG TH TRNG Nguyn Th Xuyn 8. MC LC XT NGHIM VI KHUN 1. Vi khun nhum soi .......................................................................................... 1 2. Vi khun test nhanh........................................................................................... 4 3. Vi khun nui cy v nh danh phng php thng thng........................... 7 4. Vi khun nui cy, nh danh h thng t ng............................................. 10 5. Vi khun nui cy, nh danh v khng thuc h thng t ng ................... 14 6. Vi khun khng thuc nh tnh...................................................................... 18 7. Vi khun khng thuc h thng t ng......................................................... 21 8. Vi khun khng thuc nh lng MIC (cho 1 loi khng sinh).................... 24 9. Vi khun khng sinh phi hp ........................................................................ 28 10. Vi khun k kh nui cy v nh danh ......................................................... 31 11. Vi khun khng nh..................................................................................... 34 13. Vi khun nh danh gii trnh t gene........................................................... 40 14. Vi khun khng thuc PCR........................................................................... 44 15. Vi khun khng thuc gii trnh t gene....................................................... 47 16. Vi h ng rut ........................................................................................... 51 17. AFB trc tip nhum Ziehl-Neelsen............................................................. 54 18. AFB trc tip nhum hunh quang............................................................... 58 19. Mycobacterium tuberculosis nui cy mi trng lng............................... 61 20. Mycobacterium tuberculosis nui cy mi trng c................................. 66 21. Mycobacterium tuberculosis Mantoux ......................................................... 71 22. Mycobacterium tuberculosis khng thuc hng 1 mi trng c............... 74 23. Mycobacterium tuberculosis khng thuc hng 1 mi trng lng ............. 79 24. Mycobacterium tuberculosis khng thuc hng 2 mi trng c............... 83 25. Mycobacterium tuberculosis khng thuc hng 2 mi trng lng ............. 88 26. Mycobacterium tuberculosis khng thuc PZA mi trng lng ................ 92 27. Mycobacterium tuberculosis pyrazinamidase............................................... 96 28. Mycobacterium tuberculosis nh danh v khng RMP XPERT ................. 99 29. Mycobacterium tuberculosis a khng LPA............................................... 102 30. Mycobacterium tuberculosis siu khng LPA............................................ 107 31. Mycobacterium tuberculosis PCR h thng t ng.................................. 113 9. 32. Mycobacterium tuberculosis Real-time PCR ............................................. 116 33. Mycobacterium tuberculosis Spoligotyping............................................... 120 34. Mycobacterium tuberculosis RFLP typing................................................. 125 35. NTM (Non Tuberculosis Mycobacteria) nui cy mi trng lng .......... 131 36. NTM (Non Tuberculosis Mycobacteria) nui cy mi trng c............ 136 37. NTM (Non Tuberculosis Mycobacteria) LPA............................................ 141 38. Mycobacterium leprae nhum soi .............................................................. 145 39. Mycobacterium leprae PCR ....................................................................... 148 40. Mycobacterium leprae mnh sinh thit ...................................................... 152 41. Vibrio cholerae nhum soi ......................................................................... 154 42. Vibrio cholerae nhum hunh quang ......................................................... 157 43. Vibrio cholerae nui cy, nh danh v khng thuc................................. 160 44. Vibrio cholerae PCR................................................................................... 163 45. Vibrio cholerae gii trnh t gene............................................................... 166 46. Neisseria gonorrhoeae nhum soi.............................................................. 169 47. Neisseria gonorrhoeae nui cy, nh danh v khng thuc ..................... 172 48. Neisseria gonorrhoeae PCR....................................................................... 175 49. Neisseria gonorrhoeae Real-time PCR ...................................................... 178 50. Neisseria meningitidis nhum soi............................................................... 182 51. Neisseria meningitidis nui cy, nh danh v khng thuc ...................... 185 52. Neisseria menigititdis PCR......................................................................... 188 54. Chlamydia test nhanh.................................................................................. 195 55. Chlamydia nhum hunh quang................................................................. 198 56. Chlamydia Ab min dch bn t ng........................................................ 201 57. Chlamydia PCR .......................................................................................... 205 58. Chlamydia Real-time PCR.......................................................................... 209 59. Chlamydia Real-time PCR h thng t ng ............................................. 213 60. Clostridium nui cy, nh danh................................................................. 216 61. Clostridium difficile min dch bn t ng............................................... 219 62. Clostridium difficile PCR............................................................................ 222 63. Leptospira test nhanh.................................................................................. 225 64. Mycoplasma pneumoniae Real-time PCR.................................................. 227 65. Mycoplasma hominis nui cy, nh danh v khng thuc ........................... 231 10. 66. Ricketsia Ab min dch bn t ng ........................................................... 234 67. Salmonella Widal ........................................................................................ 237 68. Streptococcus pyogenes ASO ..................................................................... 240 69. Treponema pallidum soi ti...................................................................... 243 70. Treponema pallidum nhum soi ................................................................. 245 71. Treponema pallidum RPR nh tnh v nh lng.................................... 248 72. Treponema pallidum TPHA nh tnh v nh lng................................. 251 73. Ureaplasma urealyticum nui cy, nh danh v khng thuc...................... 255 XT NGHIM VIRUS 74. Virus test nhanh........................................................................................... 258 75. Virus Ag min dch t ng........................................................................ 260 76. Virus Ab min dch t ng........................................................................ 264 77. HBsAg test nhanh ....................................................................................... 268 78. HBsAg min dch bn t ng.................................................................... 271 79. HBsAg min dch t ng........................................................................... 275 80. HBsAg khng nh...................................................................................... 279 81. HBsAg nh lng ...................................................................................... 283 82. HBsAb min dch bn t ng.................................................................... 287 83. HBsAb nh lng ...................................................................................... 291 84. HBc IgM min dch bn t ng ................................................................ 295 85. HBc IgM min dch t ng ....................................................................... 299 86. HBc total min dch bn t ng ................................................................ 303 87. HBc total min dch t ng....................................................................... 307 88. HBeAg min dch bn t ng ................................................................... 311 89. HBeAg min dch t ng .......................................................................... 315 90. HBeAb min dch bn t ng ................................................................... 318 91. HBeAb min dch t ng .......................................................................... 322 92. HBV o ti lng Real-time PCR .............................................................. 325 93. HBV o ti lng h thng t ng............................................................ 329 94. HBV genotype PCR.................................................................................... 333 95. HBV genotype Real-time PCR ................................................................... 336 96. HBV genotype gii trnh t gene ................................................................ 340 97. HBV khng thuc Real-time PCR (Cho 1 loi thuc)................................ 343 11. 98. HBV khng thuc gii trnh t gene........................................................... 346 99. HCV Ab min dch bn t ng................................................................. 349 100. HCV Ab min dch t ng...................................................................... 353 101. HCV Ag/Ab min dch bn t ng......................................................... 357 102. HCV Core Ag min dch t ng............................................................. 361 103. HCV PCR.................................................................................................. 365 104. HCV o ti lng Real-time PCR ............................................................ 368 105. HCV o ti lng h thng t ng ......................................................... 372 106. HCV genotype Real-time PCR................................................................. 375 107. HCV genotype gii trnh t gene.............................................................. 379 108. HAV IgM min dch bn t ng............................................................. 382 109. HAV IgM min dch t ng.................................................................... 386 110. HAV total min dch bn t ng............................................................. 390 111. HAV total min dch t ng ................................................................... 394 112. HDV Ag min dch bn t ng............................................................... 398 113. HDV IgM min dch bn t ng............................................................. 402 114. HDV Ab min dch bn t ng............................................................... 406 115. HEV IgM test nhanh................................................................................. 409 116. HEV IgM min dch bn t ng ............................................................. 412 117. HIV Ab test nhanh .................................................................................... 415 118. HIV Ag/Ab test nhanh.............................................................................. 418 120. HIV Ag/Ab min dch bn t ng .......................................................... 425 121. HIV Ag/Ab min dch t ng ................................................................. 429 122. HIV khng nh ........................................................................................ 433 123. HIV o ti lng Real-time PCR.............................................................. 437 124. HIV o ti lng h thng t ng........................................................... 440 125. HIV genotype gii trnh t gene ............................................................... 444 128. Dengue virus NS1Ag/IgM/IgG test nhanh ............................................... 453 129. Dengue virus IgM/IgG test nhanh ............................................................ 456 130. Dengue virus IgM min dch bn t ng................................................ 459 131. Dengue virus IgG min dch bn t ng................................................. 463 132. Dengue virus PCR .................................................................................... 467 133. Dengue virus serotype PCR...................................................................... 470 12. 134. CMV IgM min dch bn t ng............................................................. 473 135. CMV IgM min dch t ng ................................................................... 477 136. CMV IgG min dch bn t ng ............................................................. 481 137. CMV IgG min dch t ng .................................................................... 485 138. CMV PCR................................................................................................. 489 139. CMV Real-time PCR ................................................................................ 492 140. CMV o ti lng h thng t ng......................................................... 495 141. CMV Avidity ............................................................................................ 498 142. HSV 1 + 2 IgM min dch bn t ng .................................................... 502 143. HSV 1+ 2 IgG min dch bn t ng ...................................................... 506 144. HSV Real-time PCR ................................................................................. 510 145. VZV Real-time PCR ................................................................................. 514 146. EBV-VCA IgM min dch bn t ng.................................................... 518 147. EBV-VCA IgG min dch bn t ng..................................................... 522 148. EBV EA-D IgG min dch bn t ng.................................................... 526 149. EBV EB-NA IgG min dch bn t ng ................................................. 530 150. EBV PCR .................................................................................................. 534 151. EBV Real-time PCR ................................................................................. 537 152. EV71 PCR................................................................................................. 541 155. Enterovirus PCR........................................................................................ 551 156. Entrovirus genotype gii trnh t gene...................................................... 554 157. Adenovirus Real-time PCR....................................................................... 557 158. BK/JC virus PCR ...................................................................................... 561 159. HPV PCR .................................................................................................. 565 160. HPV Real-time PCR ................................................................................. 568 161. HPV Genotype Real-time PCR................................................................. 572 162. HPV genotype PCR h thng t ng ...................................................... 575 163. HPV genotype gii trnh t gene............................................................... 579 164. Influenza virus A, B test nhanh................................................................. 582 165. Influenza virus A, B Real-time PCR......................................................... 584 166. Influenza virus A, B gii trnh t gene ..................................................... 587 167. JEV IgM min dch bn t ng............................................................... 590 168. Measle virus Ab min dch bn t ng ................................................... 593 13. 169. Rotavirus test nhanh.................................................................................. 596 170. RSV Ab min dch bn t ng................................................................ 599 171. RSV Real-time PCR ................................................................................. 602 172. Rubella virus IgM min dch bn t ng................................................ 605 173. Rubella virus IgM min dch t ng....................................................... 609 174. Rubella virus IgG min dch bn t ng................................................. 613 175. Rubella virus IgG min dch t ng ....................................................... 617 176. Rubella virus PCR .................................................................................... 621 177. Rubella virus gii trnh t gene................................................................. 624 XT NGHIM K SINH TRNG - VI NM 178. Hng cu, bch cu trong phn soi ti ................................................... 627 179. Hng cu trong phn test nhanh ............................................................... 630 180. n bo ng rut soi ti..................................................................... 633 181. n bo ng rut nhum soi................................................................ 636 182. Trng giun, sn soi ti............................................................................ 639 183. Trng giun soi tp trung ........................................................................... 642 184. Strongyloides stercoralis (Giun ln) u trng soi ti........................... 645 185. Angiostrongylus cantonensis (Giun trn chut) Ab min dch bn t ng.......................................................................................................... 648 186. Clonorchis/Opisthorchis (Sn l gan nh) Ab min dch bn t ng.......................................................................................................... 651 187. Cysticercus cellulosae (Sn ln) Ab min dch bn t ng.................... 654 188. Entamoeba histolytica (Amip) Ab min dch bn t ng....................... 657 189. Fasciola (Sn l gan ln) Ab min dch bn t ng............................... 660 190. Filaria (Giun ch) u trng trong mu nhum soi.................................... 663 191. Gnathostoma(Giun u gai) Ab Min dch bn t ng.......................... 666 192. Plasmodium (K sinh trng st rt) nhum soi nh tnh ........................ 669 193. Plasmodium (K sinh trng st rt) nhum soi nh lng ..................... 672 194. Plasmodium (K sinh trng st rt) Ag test nhanh .................................. 675 195. Strongyloides stercoralis (Giun ln) Ab min dch bn t ng........... 677 196. Toxocara (Giun a ch mo) Ab min dch bn t ng....................... 680 197. Toxoplasma IgM min dch t ng......................................................... 683 198. Toxoplasma IgG min dch t ng ......................................................... 687 199. Toxoplasma Avidity min dch t ng ................................................... 691 14. 200. Demodex soi ti ...................................................................................... 695 201. Demodex nhum soi.................................................................................. 698 202. Phthirus pubis (Rn mu) soi ti.............................................................. 701 203. Phthirus pubis (Rn mu) nhum soi ......................................................... 703 204. Sarcoptes scabies hominis (Gh) soi ti................................................. 706 205. Sarcoptes scabies hominis (Gh) nhum soi ............................................ 709 206. Cysticercus cellulosae (Sn ln) u trng soi mnh sinh thit................. 712 207. Gnathostoma u trng soi mnh sinh thit ............................................... 715 208. Taenia (Sn dy) soi ti nh danh........................................................ 718 209. Toxocara (Giun a ch, mo) soi mnh sinh thit................................ 721 210. Trichinella spiralis (Giun xon) soi mnh sinh thit................................ 724 211. Trichomonas vaginalis soi toi................................................................ 727 212. Trichomonas vaginalis nhum soi............................................................ 730 213. Pneumocystis jirovecii nhum soi............................................................. 733 214. Vi nm soi ti.......................................................................................... 736 215. Vi nm test nhanh...................................................................................... 739 216. Vi nm nhum soi..................................................................................... 742 217. Vi nm nui cy v nh danh bng phng php thng thng............. 745 218. Vi nm nui cy v nh danh h thng t ng ...................................... 748 219. Vi nm nui cy, nh danh v khng thuc h thng t ng ................ 751 220. Vi nm khng nh (tham chiu)............................................................... 754 221. Vi nm khng thuc nh lng (MIC) (Cho 1 loi khng sinh) ............. 757 222. Vi nm PCR .............................................................................................. 760 223. Vi nm gii trnh t gene .......................................................................... 763 XT NGHIM NH GI NHIM KHUN BNH VIN 224. Vi sinh vt cy kim tra khng kh ........................................................... 766 225. Vi sinh vt cy kim tra bn tay................................................................ 770 226. Vi sinh vt cy kim tra dng c tit trng.......................................... 773 227. Vi sinh vt cy kim tra b mt................................................................. 776 228. Vi sinh vt cy kim tra nc sinh hot.................................................... 779 229. Vi sinh vt cy kim tra nc thi ............................................................ 783 230. Vi khun khng thuc - Pht hin ngi mang ........................................ 787 231. Vi khun gy nhim trng bnh vin - Pht hin ngun nhim ............... 841 15. CC T VIT TT BCN: Bch cu a nhn CLIA: Chemiluminescence immunoassay CLSI: Clinical and laboratory standards institute CO: Cut off DNA: Deoxyribonucleic acid ELISA: Enzyme-linked immunosorbant assay EQAS: External quality assessment scheme HBV: Hepatitis B virus HCV: Hepatitis C virus HIV: Human immunodeficiency virus HPC: High positive control HT: Huyt thanh I: Intermediate IU: International unit KKT: Khng khng th KN-KT: Khng nguyn - Khng th KST: K sinh trng LPC: Low positive control NC: Negative control OD: Optical density PC: Positive control PCR: Polymerase chain reaction QC: Quality control R: Resistant RNA: Ribonucleic acid RT PCR: Reverse transcription polymerase chain reaction S: Susceptible VD: V d VK: Vi khun VSV: Vi sinh vt 16. 1 1. Vi khun nhum soi I. MC CH V NGUYN L 1. Mc ch Nhn nh s b hnh nh vi khun v cc hnh nh t bo (nu c) trc tip t bnh phm. 2. Nguyn l nh gi hnh th, kch thc, tnh cht bt mu, cch sp xp ca vi khun v cc hnh nh t bo (nu c) bng k thut nhum v soi di knh hin vi quang hc. II. CHUN B 1. Ngi thc hin - Ngi thc hin: Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh. - Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh. 2. Phng tin, ha cht Phng tin, ha cht nh v d di y hoc tng ng. 2.1. Trang thit b - My ly tm (cn cho mt s loi bnh phm) - T an ton sinh hc cp 2 - Knh hin vi quang hc - Dng c sy lam (nu c) 2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim) STT Chi ph ha cht, vt t tiu hao n v S lng 1 L ly bnh phm L 1,000 2 Que cy Ci 1,000 3 Lam knh Ci 2,000 4 Du soi knh ml 1,000 5 Cn 96 lau knh ml 1,000 6 Nc mui sinh l ml 5,000 7 Thuc nhum fuchsin ml 5,000 8 Thuc nhum tm gentian ml 5,000 9 Cn ty 95% ml 10,000 17. 2 10 Lugol ml 5,000 11 Thuc nhum xanh methylen ml 5,000 12 Bng Kg 0,001 13 Cn 90 (v sinh dng c) ml 10,000 14 n cn Ci 0,0001 15 Panh Ci 0,0001 16 Khay ng bnh phm Ci 0,0001 17 Hp vn chuyn bnh phm Ci 0,0001 18 M Ci 0,020 19 Khu trang Ci 0,020 20 Gng tay i 3,000 21 Gng tay x l dng c i 0,020 22 Qun o bo h B 0,001 23 Acid ngm lam ml 10,000 24 ng nghim thy tinh ng 1,000 25 Bt vit knh Ci 0,020 26 Bt bi Ci 0,010 27 Bt la Ci 0,010 28 S lu kt qu xt nghim T 0,001 29 Cn st trng tay nhanh ml 1,000 30 Dung dch nc ra tay ml 8,000 31 Khn lau tay Ci 0,030 32 Giy tr kt qu xt nghim T 2,000 33 QC (nu thc hin) * 0,1 34 EQAS (nu thc hin) * 0,005 * Ghi ch: - Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng 10 mu cho 1 ln tin hnh k thut). - Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi s ln ngoi kim trung bnh 2 ln/1 nm). 3. Bnh phm 18. 3 Cc loi bnh phm c ch nh xt nghim vi khun. 4. Phiu xt nghim in y thng tin theo mu phiu yu cu. III. CC BC TIN HNH Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn. - Nh dung dch tm gentian, ph kn ni dn phin, duy tr 1 - 2 pht - dung dch tm gentian, ra tiu bn di vi nc chy nh - Nh dung dch lugol, 30 giy - dung dch lugol, ra nc - Ty mu: nh vi git cn 95% ln tiu bn, nghing i nghing li cho cn chy t cnh n sang cnh kia. Khi thy mu tm trn lam knh va phai ht th ra nc ngay. - Nh dung dch fuchsin, 1 - 2 pht - Ra nc k, kh tiu bn, soi knh hin vi. IV. NHN NH KT QU Ch c kt qu khi QC t tiu chun. 1. nh gi hnh nh vi khun trn tiu bn nhum Gram - Soi di vt knh du (x100) - Din gii v c kt qu: Vi khun Gram (+) bt mu tm sm ca gentian. Vi khun Gram (-) bt mu ca fuchsin. 2. nh gi hnh nh t bo trn tiu bn nhum n (nu c). V. NHNG SAI ST V X TR Gram dng gi: Ty cn cha thi gian Gram m gi: + Tui ca mu cy vi khun nh hng ln tnh cht nhum Gram cc mu cy vi khun thi gian qu lu. + Ty cn qu lu v trng khng k. Nhum li tiu bn khi nghi ng kt qu khng chnh xc. 19. 4 2. Vi khun test nhanh I. MC CH V NGUYN L 1. Mc ch Xc nh nhanh s c mt ca vi khun gy bnh trong bnh phm thng qua pht hin s c mt ca khng nguyn vi khun. 2. Nguyn l Pht hin khng nguyn vi khun da trn phn ng kt hp c hiu gia khng nguyn vi khng huyt thanh mu. II. CHUN B 1. Ngi thc hin - Ngi thc hin: Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh. - Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh. 2. Phng tin, ha cht Phng tin, ha cht nh v d di y hoc tng ng. 2.1. Trang thit b 2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim) STT Chi ph ha cht, vt t tiu hao n v S lng 1 Bng Kg 0,001 2 Dy gar Ci 0,001 3 Cn ml 1,000 4 Bm kim tim Ci 1,000 5 Panh Ci 0,0001 6 Khay ng bnh phm Ci 0,0001 7 Hp vn chuyn bnh phm Ci 0,001 8 Tube ng bnh phm Ci 1,000 9 Sinh phm chn on Test 1,000 10 u cn 200 ul Ci 2,000 11 Giy thm Cun 0,100 12 Giy xt nghim T 2,000 13 S lu kt qu xt nghim T 0,001 20. 5 14 Bt vit knh Ci 0,020 15 Bt bi Ci 0,010 16 M Ci 0,020 17 Khu trang Ci 0,020 18 Gng tay i 0,100 19 Gng tay x l dng c i 0,020 20 Qun o bo h B 0,005 21 Dung dch nc ra tay ml 8,000 22 Cn st trng tay nhanh ml 1,000 23 Dung dch kh trng ml 10,000 24 Khn lau tay Ci 0,010 25 QC (nu thc hin) * 0,1 26 EQAS (nu thc hin) * 0,005 Thc hin xt nghim 05 mu bnh phm/ln. * Ghi ch: - Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng 10 mu cho 1 ln tin hnh k thut). - Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi s ln ngoi kim trung bnh 2 ln/1 nm). 3. Bnh phm Cc loi bnh phm ch nh lm test nhanh. 4. Phiu xt nghim in y thng tin theo mu phiu yu cu III. CC BC THC HIN Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn. 1. Ly bnh phm Theo ng quy nh ca chuyn ngnh Vi sinh (Xem Ph lc 1). 2. Tin hnh k thut B sinh phm Slidex Meningite Kit 5 (VD) pht hin 5 cn nguyn vi khun gy vim mng no. 21. 6 Cc bc thc hin K thut 1. a sinh phm ra ngoi nhit phng nh s th t cc bnh phm v s th t trn tm knh 2. Lc nh nhng l cha ht latex, khng c lc qu mnh 3. Dng pipet nh 1 git dch no ty ca ngi bnh vo tng ng nh s trn phin knh. Nh chng dng v chng m 4. Lc nh nhng l c cha ht latex v nh mt git vo cnh git dch no ty ca ngi bnh 5. Trn u 2 loi vi nhau bng que trn ph u b mt ca mi . 6. Lc u c phin knh bng tay hoc dng my lc 80-100 vng trong 10 pht 7. c kt qu IV. NHN NH KT QU - chng dng: c nhin tng ngng kt ht latex - chng m: khng c nhin tng ngng kt, hn dch nhn thy mn, ng nht - khng c hin tng ngng kt, hn dch nhn thy mn, ng nht: m tnh - c hin tng ngng kt, hn dch nhn thy th, c ht ngng kt r trn nn en. V. NHNG SAI ST V X TR Lng bnh phm a vo qu nhiu hoc qu t u c th nh hng n kt qu. Do vy, phi tin hnh ly th tch bnh phm ng theo yu cu (Xem Ph lc 6). 22. 7 3. Vi khun nui cy v nh danh phng php thng thng I. MC CH V NGUYN L 1. Mc ch Pht hin v nh danh vi khun gy bnh bng phng php nui cy kinh in. 2. Nguyn l Vi khun c nh danh da vo c im nui cy, mt s tnh cht chuyn ha, cc c im v hnh thi hc v c th kt hp vi tnh cht khng nguyn. II. CHUN B 1. Ngi thc hin - Ngi thc hin: Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh. - Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh. 3. Phng tin, ha cht Phng tin, ha cht nh v d di y hoc tng ng. 2.1.Trang thit b - T an ton sinh hc cp 2 - Knh hin vi quang hc - T m thng - T m CO2 - My tnh ci phn mm c API (nu c) 2.2.Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim) STT Chi ph ha cht, vt t tiu hao n v S lng 1 L ly bnh phm L 1,000 2 Que cy Ci 1,000 3 Lam knh Ci 2,000 4 Du soi knh ml 1,000 5 Cn 96 lau knh ml 1,000 6 Nc mui sinh l ml 5,000 23. 8 7 Thuc nhum fuchsin ml 5,000 8 Thuc nhum tm gentian ml 5,000 9 Cn ty 95 ml 10,000 10 Lugol ml 5,000 11 Thuc nhum xanh methylen ml 5,000 12 Mi trng nui cy a 1,500 13 B sinh vt ha hc (b API ) Test 0,500 14 Bng Kg 0,001 15 Cn 90 (v sinh dng c) ml 10,000 16 n cn Ci 0,0001 17 Panh Ci 0,0001 18 Khay ng bnh phm Ci 0,0001 19 Hp vn chuyn bnh phm Ci 0,0001 20 M Ci 0,020 21 Khu trang Ci 0,020 22 Gng tay i 3,000 23 Gng tay x l dng c i 0,020 24 Qun o bo h B 0,001 25 Acid ngm lam ml 10,000 26 ng nghim thy tinh ng 1,000 27 Bt vit knh Ci 0,020 28 Bt bi Ci 0,010 29 Bt la Ci 0,010 30 S lu kt qu xt nghim T 0,001 31 Cn st trng tay nhanh ml 1,000 32 Dung dch nc ra tay ml 8,000 33 Khn lau tay Ci 0,030 34 Giy tr kt qu xt nghim T 2,000 35 QC (nu thc hin) * 0,1 36 EQAS (nu thc hin) * 0,005 Mi trng nui cy v ha cht nh danh vi khun c tnh trn t l dng tnh trung bnh l 50 % cho cc loi bnh phm. 24. 9 * Ghi ch: - Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng 10 mu cho 1 ln tin hnh k thut). - Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi s ln ngoi kim trung bnh 2 ln/1 nm). 3. Bnh phm Tt c cc loi bnh phm c ch nh nui cy tm vi khun gy bnh. 4. Phiu xt nghim in y thng tin theo mu phiu yu cu III. CC BC TIN HNH Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn. 1. Ly bnh phm Theo ng quy nh ca chuyn ngnh Vi sinh (Xem Ph lc 1). 2. Tin hnh k thut - Nhum soi bnh phm, nh gi s b - Nui cy bnh phm vo mi trng phn lp - m qua m - Bt khun lc nghi ng - Nhum soi, th nghim cc th nghim sinh vt ha hc n gin v nh danh bng cc b sinh vt ha hc (b API ) IV. NHN NH KT QU - Dng tnh: Phn lp v nh danh c vi khun gy bnh. Tr kt qu tn vi khun n mc chi v/hoc loi. - m tnh: Khng tm thy hoc khng phn lp c vi khun gy bnh. V. NHNG SAI ST V X TR Quy trnh ny ch p dng tm vi khun hiu k kh ty tin d nui cy, khng p dng cho cc vi khun k kh bt buc. Kt qu m tnh khng c ngha l khng c vi khun gy bnh trong bnh phm m l khng tm thy cn nguyn vi khun gy bnh c th phn lp c bng quy trnh nui cy ny. Nu c yu cu tm cn nguyn vi khun him gp, phi ghi yu cu c th. Bnh phm ly, vn chuyn v bo qun khng ng yu cu c th a n kt qu m tnh hoc dng tnh gi (Xem Ph lc 6). 25. 10 4. Vi khun nui cy, nh danh h thng t ng I. MC CH V NGUYN L 1. Mc ch Pht hin vi khun gy bnh bng phng php nui cy, nh danh trn h thng t ng. 2. Nguyn l Vi khun c nh danh bng my t ng da trn mt s tnh cht chuyn ha kt hp vi cc c im v hnh thi hc v tnh cht bt mu. II. CHUN B 1. Ngi thc hin - Ngi thc hin Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh. - Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh. 2. Phng tin, ha cht Phng tin, ha cht nh v d di y hoc tng ng. 2.1. Trang thit b - Knh hin vi quang hc - T m thng - T m CO2 - H thng nh danh t ng - My o c - L hp t - T an ton sinh hc cp 2 2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim) STT Chi ph ha cht, vt t tiu hao n v S lng 1 L ly bnh phm L 1,000 2 Que cy Ci 2,000 3 Lam knh Ci 2,000 4 Du soi knh ml 1,000 5 Cn 96 lau knh ml 1,000 6 Nc mui sinh l ml 5,000 26. 11 7 Thuc nhum fuchsin ml 5,000 8 Thuc nhum tm gentian ml 5,000 9 Cn ty 95 ml 10,000 10 Lugol ml 5,000 11 Thuc nhum xanh methylen ml 5,000 12 Mi trng nui cy a 1,500 13 Panel nh danh theo h thng my t ng Test 0,500 14 B Panel chng B 0,0002 15 Bm kim tim Ci 1,000 16 Bng Kg 0,001 17 Cn 90 (v sinh dng c) ml 10,000 18 n cn Ci 0,0001 19 Panh Ci 0,0001 20 Khay ng bnh phm Ci 0,0001 21 Hp vn chuyn bnh phm Ci 0,0001 22 M Ci 0,020 23 Khu trang Ci 0,020 24 Gng tay i 3,000 25 Gng tay x l dng c i 0,020 26 Qun o bo h B 0,001 27 Acid ngm lam ml 10,000 28 ng nghim thy tinh ng 1,000 29 Bt vit knh Ci 0,020 30 Bt bi Ci 0,010 31 Bt la Ci 0,010 32 S lu kt qu xt nghim T 0,001 33 Cn st trng tay nhanh ml 1,000 34 Dung dch nc ra tay ml 8,000 35 Khn lau tay Ci 0,030 36 Giy tr kt qu xt nghim T 2,000 37 QC (nu thc hin) * 0,1 38 EQAS (nu thc hin) * 0,005 27. 12 Mi trng nui cy c tnh trn t l dng l 50 % so vi tng s bnh phm gi xt nghim. B Panel s dng kim chun c tnh trn 5000 Test/1 nm. * Ghi ch: - Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng 10 mu cho 1 ln tin hnh k thut). - Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi s ln ngoi kim trung bnh 2 ln/1 nm). 3. Bnh phm Tt c cc loi bnh phm c ch nh nui cy tm vi khun gy bnh. 4. Phiu xt nghim in y thng tin theo mu phiu yu cu. II. CC BC TIN HNH Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn. 1. Ly bnh phm Theo ng quy nh ca chuyn ngnh Vi sinh (Xem ph lc 1). 2. Tin hnh k thut - Nhum soi bnh phm, nh gi s b - Nui cy bnh phm vo mi trng phn lp - m qua m - Bt khun lc nghi ng - Nhum soi, th nghim cc tnh cht sinh vt ha hc n gin v nh danh bng h thng t ng. IV. NHN NH KT QU - Dng tnh: Phn lp v nh danh c vi khun gy bnh. Tr kt qu tn vi khun n mc chi v/hoc loi. - m tnh: Khng tm thy hoc khng phn lp c vi khun gy bnh. V. NHNG SAI ST V X TR Quy trnh ny ch p dng tm vi khun hiu k kh ty tin d nui cy, khng p dng cho cc vi khun k kh bt buc. Kt qu m tnh khng c ngha l khng c vi khun gy bnh trong bnh phm m l khng tm thy cn nguyn vi khun gy bnh c th phn lp c bng quy trnh nui cy ny. Nu c yu cu tm cn nguyn vi khun no , phi ghi yu cu c th. 28. 13 Bnh phm ly, vn chuyn v bo qun khng ng yu cu c th a n kt qu m tnh hoc dng tnh gi (Xem Ph lc 6). 29. 14 5. Vi khun nui cy, nh danh v khng thuc h thng t ng I. MC CH V NGUYN L 1. Mc ch Pht hin vi khun gy bnh bng phng php nui cy, nh danh v khng thuc trn h thng t ng. 2. Nguyn l Vi khun c nh danh bng my t ng da trn mt s tnh cht chuyn ha kt hp vi cc c im v hnh thi hc v tnh cht bt mu; v xc nh mc nhy cm vi khng sinh bng phng php khng sinh pha long. II. CHUN B 1. Ngi thc hin - Ngi thc hin Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh. - Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh. 2. Phng tin, ha cht Phng tin, ha cht nh v d di y hoc tng ng. 2.1. Trang thit b - Knh hin vi quang hc - T m thng - T m CO2 - H thng nh danh t ng - My o c - L hp t - T an ton sinh hc cp 2 2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim) STT Chi ph ha cht, vt t tiu hao n v S lng 1 L ly bnh phm L 1,000 2 Lam knh Ci 2,000 3 Que cy Ci 2,000 4 Du soi knh ml 1,000 30. 15 5 Cn 96 lau knh ml 1,000 6 Nc mui sinh l ml 5,000 7 Thuc nhum Fuchsin ml 5,000 8 Thuc nhum tm Gentian ml 5,000 9 Cn ty 95 ml 10,000 10 Lugol ml 5,000 11 Thuc nhum xanh methylen ml 5,000 12 Mi trng nui cy a 1,500 13 Panel nh danh theo h thng my t ng Test 1,000 14 B Panel chng B 0,0002 15 Bm kim tim Ci 1,000 16 Bng Kg 0,001 17 Cn 90 (v sinh dng c) ml 10,000 18 n cn Ci 0,0001 19 Panh Ci 0,0001 20 Khay ng bnh phm Ci 0,0001 21 Hp vn chuyn bnh phm Ci 0,0001 22 M Ci 0,020 23 Khu trang Ci 0,020 24 Gng tay i 3,000 25 Gng tay x l dng c i 0,020 26 Qun o bo h B 0,001 27 Acid ngm lam ml 10,000 28 ng nghim thy tinh ng 1,000 29 Bt vit knh Ci 0,020 30 Bt bi Ci 0,010 31 Bt la Ci 0,010 32 S lu kt qu xt nghim T 0,001 33 Cn st trng tay nhanh ml 1,000 34 Dung dch nc ra tay ml 8,000 35 Khn lau tay Ci 0,030 36 Giy tr kt qu xt nghim T 2,000 31. 16 37 QC (nu thc hin) * 0,1 38 EQAS (nu thc hin) * 0,005 Mi trng nui cy c tnh trn t l dng l 50 % so vi tng s bnh phm gi xt nghim. B Panel s dng kim chun c tnh trn 5000 Test/1 nm. * Ghi ch: - Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng 10 mu cho 1 ln tin hnh k thut). - Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi s ln ngoi kim trung bnh 2 ln/1 nm). 3. Bnh phm Tt c cc loi bnh phm c ch nh nui cy tm vi khun gy bnh. 4. Phiu xt nghim in y thng tin theo mu phiu yu cu. III. CC BC TIN HNH Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn. 1. Ly bnh phm Theo ng quy nh ca chuyn ngnh Vi sinh (Xem ph lc 1). 2. Tin hnh k thut - Nhum soi bnh phm, nh gi s b - Nui cy bnh phm vo mi trng phn lp - m qua m - Bt khun lc nghi ng - Nhum soi, th nghim cc tnh cht sinh vt ha hc n gin v nh danh bng h thng t ng. IV. NHN NH KT QU - Dng tnh: Phn lp v nh danh c vi khun gy bnh. Tr kt qu tn vi khun n mc chi v/hoc loi. - m tnh: Khng tm thy hoc khng phn lp c vi khun gy bnh. V. NHNG SAI ST V X TR Quy trnh ny ch p dng tm vi khun hiu k kh ty tin d nui cy, khng p dng cho cc vi khun k kh bt buc. Kt qu m tnh khng c 32. 17 ngha l khng c vi khun gy bnh trong bnh phm m l khng tm thy cn nguyn vi khun gy bnh c th phn lp c bng quy trnh nui cy ny. S dng sai panel nh danh cho loi vi khun cn xc nh. Cn chn la chnh xc panel nh danh cho tng loi vi khun. Nu c yu cu tm cn nguyn vi khun no , phi ghi yu cu c th. Bnh phm ly, vn chuyn v bo qun khng ng yu cu c th a n kt qu m tnh hoc dng tnh gi (Xem Ph lc 6). 33. 18 6. Vi khun khng thuc nh tnh I. MC CH V NGUYN L 1. Mc ch Xc nh mc nhy cm vi khng sinh ca cc chng vi khun gy bnh. 2. Nguyn l Mc nhy cm vi khng sinh ca cc chng vi khun th nghim c nh gi bng phng php khng sinh khuch tn trong thch. S pht trin ca vi khun s b c ch khi khng sinh t n mt nng nht nh. ng knh vng c ch t l thun vi mc nhy cm c phin gii ra cc phn loi S (sensitive nhy cm), I (intermediate trung gian), hoc R (resistant khng) khi so snh vi bng chun CSLI cp nht hng nm. II. CHUN B 1. Ngi thc hin - Ngi thc hin Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh. - Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh. 2. Phng tin, ha cht Phng tin, ha cht nh v d di y hoc tng ng. 2.1. Trang thit b - T m - T an ton sinh hc cp 2 - My lc - ng c chun McFarland 0.5 2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim) STT Chi ph ha cht, vt t tiu hao n v S lng 1 Khoanh giy khng sinh Khoanh 22,000 2 Que cy Ci 1,000 3 Thch Muller Hinton a 3,000 4 Thch mu a 1,000 5 Giy khng sinh T 2,000 6 Bng Kg 0,001 7 Cn 90 (v sinh dng c) ml 10,000 34. 19 8 Panh Ci 0,0001 9 Khay ng bnh phm Ci 0,0001 10 Hp vn chuyn bnh phm Ci 0,0001 11 M Ci 0,050 12 Khu trang Ci 0,050 13 Gng tay i 3,000 14 Gng tay x l dng c i 0,030 15 Qun o bo h B 0,001 16 Nc mui sinh l ml 5,000 17 ng nghim thy tinh ng 1,000 18 Tm bng v trng Ci 1,000 19 Bt vit knh Ci 0,020 20 Bt bi Ci 0,010 21 Bt la Ci 0,010 22 S lu kt qu xt nghim Quyn 0,001 23 Cn st trng tay nhanh ml 1,000 24 Dung dch nc ra tay ml 8,000 25 Khn lau tay Ci 0,050 26 QC (nu thc hin) * 0,1 27 EQAS (nu thc hin) * 0,005 * Ghi ch: - Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng 10 mu cho 1 ln tin hnh k thut). - Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi s ln ngoi kim trung bnh 2 ln/1 nm). 3. Bnh phm Chng vi khun c xc nh l cn nguyn gy bnh, thun v mi. 4. Phiu xt nghim in y thng tin theo mu phiu yu cu. III. CC BC TIN HNH Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn. 35. 20 1. Ly bnh phm Chng vi khun cn th nghim c nui cy thun nht trong iu kin ti u v ang giai on pht trin mnh (nui cy sau 16-24 gi). 2. Tin hnh k thut 2.1. Pha huyn dch vi khun Dng que cy ly vi khun t 3-5 khun lc c hnh thi ging nhau nghin u vo ng nc mui sinh l 5 ml, lc u trn my lc c huyn dch ng nht. So snh c ca huyn dch vi khun vi c ca ng McFarland 0,5. 2.2. Dn u canh khun ln mt a thch Dng tm bng v trng nhng vo ng huyn dch vi khun pha trn, p nh v xoay trn tm bng trn thnh bn ca ng huyn dch vi khun loi bt phn huyn dch vi khun thm vo u tm bng. Sau , ria u que tm bng trn ton b mt a thch Mueller-Hinton sao cho vi khun c dn u ln trn ton b b mt a thch 2.3. t cc khoanh giy khng sinh ln mt thch Cc khoanh giy sau khi t cn c n xung va phi m bo chng c tip xc hon ton vi mt thch. Trong vng 15 pht sau khi t khoanh giy khng sinh, cc a thch phi c lt p trong t m 35C. 2.4. a thch trong t m 16 - 24 gi IV. NHN NH KT QU Ch c kt qu khng sinh chng ngi bnh khi kt qu QC t. o ng knh vng c ch (bao gm c ng knh ca khoanh giy khng sinh) tnh theo mm. Phin gii ng knh vng c ch ra kt qu S, I, R theo hng dn ca CLSI cp nht hng nm. V. NHNG SAI ST V X TR Quy trnh ny ch p dng cho cc chng vi khun hiu k kh ty tin d nui cy, khng p dng cho cc vi khun k kh bt buc. Sai st c th gp khi: - Ln hai hay nhiu chng vi khun - Vi khun mc qu dy hoc qu tha. Phi tin hnh lm li khi thy cc hin tng nh trn. 36. 21 7. Vi khun khng thuc h thng t ng I. MC CH V NGUYN L 1. Mc ch Xc nh mc nhy cm vi khng sinh ca cc chng vi khun gy bnh. 2. Nguyn l Mc nhy cm vi khng sinh ca cc chng vi khun th nghim c nh gi da trn gi tr ca nng c ch ti thiu - MIC. Gi tr MIC s c phin gii ra phn loi S (sensitive nhy cm), I (intermediate trung gian), hoc R (resistant khng) nh h thng t ng. II. CHUN B 1. Ngi thc hin - Ngi thc hin: Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh. - Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh. 2. Phng tin, ha cht Phng tin, ha cht nh v d di y hoc tng ng. 2.1. Trang thit b - My nh danh, khng sinh t ng (v d H thng Vitek 2 Compact) - T an ton sinh hc cp 2 - My vi tnh - My in 2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim) STT Chi ph ha cht, vt t tiu hao n v S lng 1 Th khng sinh Th 1,000 2 Que cy Ci 1,000 3 Blood agar base 500G ml 75,000 4 Dng c ly bnh phm L 1,000 5 Mac Conkey agar ml 75,000 6 Khay ng bnh phm Ci 0,0001 7 M Ci 0,030 8 Khu trang Ci 0,030 37. 22 9 Gng tay i 3,000 10 Gng tay x l dng c i 0,030 11 Qun o bo h Ci 0.0005 12 ng nghim thy tinh ng 1,000 13 Dung dch nc ra tay ml 8,000 14 Khn lau tay Ci 0,030 15 Giy tr kt qu xt nghim T 4,000 16 Presept Vin 1,000 17 QC (nu thc hin) * 0,1 18 EQAS (nu thc hin) * 0,005 * Ghi ch: - Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng 10 mu cho 1 ln tin hnh k thut). - Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi s ln ngoi kim trung bnh 2 ln/1 nm). 3. Bnh phm Chng vi khun c xc nh l cn nguyn gy bnh, thun v mi. 4. Phiu xt nghim in y thng tin theo mu phiu yu cu III. CC BC TIN HNH Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn. 1. Ly bnh phm Chng vi khun cn th nghim c nui cy thun nht trong iu kin ti u v ang giai on pht trin mnh (nui cy sau 16-24 gi). 2. Tin hnh k thut - Chun b mu - Np ng nghim v card test ng loi vo cassette - Nhp mu vo my - Khai bo thng tin - To thng tin bnh nhn - Xem kt qu ca mu 38. 23 IV. NHN NH KT QU Ch c kt qu khng sinh chng ngi bnh khi kt qu QC t. Ghi nhn cc kt qu S, I hay R cng vi gi tr MIC. V. NHNG SAI ST V X TR Quy trnh ny ch p dng cho cc chng vi khun hiu k kh ty tin d nui cy, khng p dng cho cc vi khun k kh bt buc. Sai st c th gp khi: - Ln hai hay nhiu chng vi khun - Chn th lm khng sinh khng ph hp vi chng vi khun cn th nghim - Chng vi khun sau khi pha khng c a vo my trong vng 30 pht Phi tin hnh lm li khi thy cc hin tng nh trn. 39. 24 8. Vi khun khng thuc nh lng MIC (cho 1 loi khng sinh) I. MC CH V NGUYN L 1. Mc ch Xc nh mc nhy cm v nng c ch ti thiu ca khng sinh vi chng vi khun gy bnh. 2. Nguyn l Etest l thanh plastic kch thc 5 x 57 mm c phn b di MIC theo g/mL v c m code tn khng sinh. C 15 nng khng sinh c pha long bc 2 gn c nh mt mt ca bng plastic. Khi bng Etest c t ln mt thch dn vi khun, khng sinh c bc nng nhanh chng khuch tn trong thch. Sau khi nui cy qua m, s xut hin vng c ch hnh elip i xng qua thanh plastic. Gi tr MIC c xc nh trc tip ti im ct ca hnh elip vi thanh Etest. Gi tr MIC s c phin gii ra phn loi S (sensitive nhy cm), I (intermediate trung gian), hoc R (resistant khng) khi so snh vi bng chun CLSI cp nht hng nm. II. CHUN B 1. Ngi thc hin - Ngi thc hin Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh. - Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh. 2. Phng tin, ha cht Phng tin, ha cht nh v d di y hoc tng ng. 2.1. Trang thit b - T m - T an ton sinh hc cp 2 - My lc - ng c chun McFarland 0.5 2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim) STT Chi ph ha cht, vt t tiu hao n v S lng 1 Thanh giy khng sinh Ci 1,000 2 Que cy Ci 1,000 3 Thch Muller Hinton a 1,000 4 Thch mu a 1,000 40. 25 5 Giy khng sinh T 2,000 6 Bng Kg 0,001 7 Cn 90 (v sinh dng c) ml 10,000 8 Panh Ci 0,0001 9 Khay ng bnh phm Ci 0,0001 10 Hp vn chuyn bnh phm Ci 0,0001 11 M Ci 0,050 12 Khu trang Ci 0,050 13 Gng tay i 3,000 14 Gng tay x l dng c i 0,030 15 Qun o bo h B 0,001 16 Nc mui sinh l ml 5,000 17 ng nghim thy tinh ng 1,000 18 Tm bng v trng Ci 1,000 19 Bt vit knh Ci 0,020 20 Bt bi Ci 0,010 21 Bt la Ci 0,010 22 S lu kt qu xt nghim Quyn 0,001 23 Cn st trng tay nhanh ml 1,000 24 Dung dch nc ra tay ml 8,000 25 Khn lau tay Ci 0,050 26 QC (nu thc hin) * 0,1 27 EQAS (nu thc hin) * 0,005 * Ghi ch: - Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng 10 mu cho 1 ln tin hnh k thut). - Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi s ln ngoi kim trung bnh 2 ln/1 nm). 3 Bnh phm Chng vi khun c xc nh l cn nguyn gy bnh, thun v mi. 41. 26 4 Phiu xt nghim in y thng tin theo mu phiu yu cu. III. CC BC TIN HNH Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn. 1. Ly bnh phm Chng vi khun cn th nghim c nui cy thun nht trong iu kin ti u v ang giai on pht trin mnh (nui cy sau 16-24 gi). 2. Tin hnh k thut 2.1. Pha huyn dch vi khun Dng que cy ly vi khun t 3-5 khun lc c hnh thi ging nhau nghin u vo ng nc mui sinh l 5 ml, lc u trn my lc c huyn dch ng nht. So snh c ca huyn dch vi khun vi c ca ng McFarland 0,5. 2.2. Dn u canh khun ln mt a thch Dng tm bng v trng nhng vo ng huyn dch vi khun pha trn, p nh v xoay trn tm bng trn thnh bn ca ng huyn dch vi khun loi bt phn huyn dch vi khun thm vo u tm bng. Sau , ria u que tm bng trn ton b mt a thch Mueller-Hinton sao cho vi khun c dn u ln trn ton b b mt a thch 2.3. t cc khoanh giy khng sinh ln mt thch t thanh Etest ln mt thch sao cho mt c ghi di nng hng ln trn v phi m bo ton b b mt ca thanh Etest c tip xc hon ton vi mt thch. Khi t xong thanh Etest khng c dch chuyn thanh Etest khi v tr. 2.4. a thch trong t m 16 - 24 gi IV. NHN NH KT QU 1. Ch c kt qu khng sinh chng ngi bnh khi kt qu QC t. 2. c kt qu Sau m 16 24 h v khi thy r vi khun mc, c gi tr MIC im ct ca hnh elip vi thanh E-test. Lm trn gi tr MIC im gia hai bc pha long ln gi tr cao hn mt bc trc khi phin gii kt qu. Phin gii kt qu MIC ra gi tr S, I hoc R theo ti liu ca CLSI cp nht hng nm. Vi cc khng sinh dit khun, phi c gi tr MIC ti im vi khun b c ch hon ton. Vi cc khng sinh km khun, phi c gi tr MIC ti im vi khun b c ch 80%. V. NHNG SAI ST V X TR 42. 27 Quy trnh ny ch p dng cho cc chng vi khun hiu k kh ty tin d nui cy, khng p dng cho cc vi khun k kh bt buc. Sai st c th gp khi: - Ln hai hay nhiu chng vi khun - Vi khun mc qu dy hoc qu tha. Phi tin hnh lm li khi thy cc hin tng nh trn. 43. 28 9. Vi khun khng sinh phi hp I. MC CH V NGUYN L 1. Mc ch Tm cp khng sinh phi hp c tc dng hip ng trn chng vi khun gy bnh. 2. Nguyn l Xc nh tc dng hip ng, i khng hoc khng khc bit (gi tr MIC) khi th nghim phi hp 2 khng sinh khc nhau vi vi khun gy bnh bng thanh E-test da trn ch s FIC (Fractional Inhibitory Concentration). II. CHUN B 1. Ngi thc hin - Ngi thc hin Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh. - Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh. 2. Phng tin, ha cht Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn. 2.1. Trang thit b - T m - T an ton sinh hc cp 2 - My lc - ng c chun McFarland 0.5 2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim) STT Chi ph ha cht, vt t tiu hao n v S lng 1 Thanh giy khng sinh Ci 4,000 2 Thch Muller Hinton a 3,000 3 Que cy Ci 1,000 4 Thch mu a 2,000 5 Giy khng sinh T 2,000 6 Bng Kg 0,001 7 Cn 90 (v sinh dng c) ml 10,000 8 Panh Ci 0,0001 9 Khay ng bnh phm Ci 0,0001 44. 29 10 Hp vn chuyn bnh phm Ci 0,0001 11 M Ci 0,050 12 Khu trang Ci 0,050 13 Gng tay i 3,000 14 Gng tay x l dng c i 0,030 15 Qun o bo h bo h B 0,001 16 Nc mui sinh l ml 5,000 17 ng nghim thy tinh ng 1,000 18 Tm bng v trng Ci 1,000 19 Bt vit knh Ci 0,020 20 Bt bi Ci 0,010 21 Bt la Ci 0,010 22 S lu kt qu xt nghim Quyn 0,001 23 Cn st trng tay nhanh ml 1,000 24 Dung dch nc ra tay ml 8,000 25 Khn lau tay Ci 0,050 26 QC (nu thc hin) * 0,1 27 EQAS (nu thc hin) * 0,005 * Ghi ch: - Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng 10 mu cho 1 ln tin hnh k thut). - Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi s ln ngoi kim trung bnh 2 ln/1 nm). 3. Bnh phm Chng vi khun c xc nh l cn nguyn gy bnh, thun v mi. 4. Phiu xt nghim in y thng tin theo mu phiu yu cu. III. CC BC TIN HNH Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn. - Pha huyn dch vi khun c c tng ng vi c McFarland 0.5 45. 30 - Dn u canh khun ln mt 2 a thch Mueller-Hinton v cy thun trn 1 a thch mu - t 2 thanh E-test ln 2 a thch dn u vi khun - cc a thch trong t m 16 - 18 gi - c kt qu gi tr MIC ca c hai khng sinh - Pha huyn dch vi khun c c tng ng vi c McFarland 0.5 ri dn u canh khun ln mt 1 a thch Mueller-Hinton v cy thun trn 1 a thch mu - t 2 thanh E-test vung gc vi nhau, giao nhau ti gi tr MIC ca mi thanh c bc 5 ln a thch dn u vi khun - cc a thch trong t m 16 - 18 gi - c kt qu gi tr MIC ca c hai khng sinh IV. NHN NH KT QU Ch c kt qu khng sinh chng ngi bnh khi kt qu QC t. c kt qu: - Tnh ch s FIC (Fractional Inhibitory Concentration) FIC ca khng sinh A = MIC ca khng sinh A trong th nghim phi hp/MIC ca khng sinh A th nghim ring FIC ca khng sinh B = MIC ca khng sinh B trong th nghim phi hp/MIC ca khng sinh B th nghim ring Ch s FIC = FIC ca khng sinh A/ FIC ca khng sinh B - Phin gii kt qu ch s FIC o Hai khng sinh c tc dng hip ng nu ch s FIC 0.5 o Hai khng sinh c tc dng i khng nu ch s FIC > 4 o Hai khng sinh phi hp khng c tc dng khc bit nu ch s FIC 4 v > 0.5 V. NHNG SAI ST V X TR Quy trnh ny ch p dng cho cc chng vi khun hiu k kh ty tin d nui cy, khng p dng cho cc vi khun k kh bt buc. Nhng trng hp tc dng i khng yu c th khng pht hin c v vng c ch nm di vng bt cho ca hai thanh E-test. Sai st c th gp khi: - Ln hai hay nhiu chng vi khun - Vi khun mc qu dy hoc qu tha. - Phi tin hnh lm li khi thy cc hin tng nh trn. 46. 31 10. Vi khun k kh nui cy v nh danh I. MC CH V NGUYN L 1. Mc ch Pht hin v nh danh vi khun k kh gy bnh bng phng php nui cy kinh in. 2. Nguyn l Vi khun c nh danh da vo c im nui cy, mt s tnh cht chuyn ha, cc c im v hnh thi hc v c th kt hp vi tnh cht khng nguyn. II. CHUN B 1. Ngi thc hin - Ngi thc hin: Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh. - Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh. 2. Phng tin, ha cht Phng tin, ha cht nh v d di y hoc tng ng. 2.1. Trang thit b - Knh hin vi quang hc - T an ton sinh hc cp 2 - H thng my nui cy vi khun k kh 2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim) STT Chi ph ha cht, vt t tiu hao n v S lng 1 L ly bnh phm L 1,000 2 Lam knh Ci 2,000 3 Que cy Ci 2,000 4 Du soi knh ml 1,000 5 Cn 96 lau knh ml 1,000 6 Nc mui sinh l ml 5,000 7 Thuc nhum fuchsin ml 5,000 8 Thuc nhum tm gentian ml 5,000 9 Cn ty 95 ml 10,000 10 Lugol ml 5,000 47. 32 11 Thuc nhum xanh methylen ml 5,000 12 Mi trng nui cy k kh chai 1,000 13 Mi trng thch mu thng a 2,000 14 Mi trng thch mu k kh a 2,000 15 B gi ng API Test 1,000 16 Bnh kh trn (tnh theo gi) Gi 168,000 17 Bm kim tim Ci 1,000 18 Bng Kg 0,001 19 Cn 90 (v sinh dng c) ml 10,000 20 n cn Ci 0,0001 21 Panh Ci 0,0001 22 Khay ng bnh phm Ci 0,0001 23 Hp vn chuyn bnh phm Ci 0,0001 24 M Ci 0,020 25 Khu trang Ci 0,020 26 Gng tay i 3,000 27 Gng tay x l dng c i 0,020 28 Qun o bo h bo h B 0,001 29 Acid ngm lam ml 10,000 30 ng nghim thy tinh ng 1,000 31 Bt vit knh Ci 0,020 32 Bt bi Ci 0,010 33 Bt la Ci 0,010 34 S lu kt qu xt nghim T 0,001 35 Cn st trng tay nhanh ml 1,000 36 Dung dch nc ra tay ml 8,000 37 Khn lau tay Ci 0,030 38 Giy tr kt qu xt nghim T 2,000 39 QC (nu thc hin) * 0,1 39 EQAS (nu thc hin) * 0,005 * Ghi ch: 48. 33 - Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng 10 mu cho 1 ln tin hnh k thut). - Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi s ln ngoi kim trung bnh 2 ln/1 nm). 3. Bnh phm Tt c cc loi bnh phm c ch nh nui cy tm vi khun k kh gy bnh. 4. Phiu xt nghim in y thng tin theo mu phiu yu cu III. CC BC TIN HNH Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn. 1. Ly bnh phm Theo ng quy nh ca chuyn ngnh Vi sinh (Xem Ph lc 1). 2. Tin hnh k thut - Nhum soi bnh phm, nh gi s b - Nui cy bnh phm vo mi trng phn lp - m qua m - Bt khun lc nghi ng - Nhum soi, th nghim cc th nghim sinh vt ha hc n gin v nh danh bng cc b sinh vt ha hc chuyn dng. - Tt c cc bc tin hnh v m u c thc hin bn trong h thng my nui cy vi khun k kh nhm m bo iu kin mi trng k kh. IV. NHN NH KT QU - Dng tnh: Phn lp v nh danh c vi khun gy bnh. Tr kt qu tn vi khun n mc chi v/hoc loi. - m tnh: Khng tm thy hoc khng phn lp c vi khun gy bnh. V. NHNG SAI ST V X TR Quy trnh ny ch p dng tm vi khun k kh bt buc v k kh ty tin. Kt qu m tnh khng c ngha l khng c vi khun gy bnh trong bnh phm m l khng tm thy cn nguyn vi khun gy bnh c th phn lp c bng quy trnh nui cy ny. Tt c cc thao tc k thut phi din ra bn trong h thng my to mi trng k kh m bo iu kin nui cy thch hp cho vi khun k kh mc. Kh trng s dng nui cy vi khun phi m bo ng nng . Nu c yu cu tm cn nguyn vi khun no , phi ghi yu cu c th. 49. 34 11. Vi khun khng nh I. MC CH V NGUYN L 1. Mc ch Pht hin vi khun gy bnh bng phng php nui cy, nh danh trn h thng t ng. 2. Nguyn l Vi khun c nh danh bng my t ng da trn mt s tnh cht chuyn ha kt hp vi cc c im v hnh thi hc v tnh cht bt mu. II. CHUN B 1. Ngi thc hin - Ngi thc hin Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh. - Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh. 2. Phng tin, ha cht Phng tin, ha cht nh v d di y hoc tng ng. 2.1. Trang thit b - Knh hin vi quang hc - T m thng - T m CO2 - H thng nh danh t ng - My o c - L hp t - T an ton sinh hc cp 2 2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim) STT Chi ph ha cht, vt t tiu hao n v S lng 1 L ly bnh phm L 1,000 2 Lam knh Ci 2,000 3 Que cy Ci 1,000 4 Du soi knh ml 1,000 5 Cn 96 lau knh ml 1,000 6 Nc mui sinh l ml 5,000 50. 35 7 Thuc nhum Fuchsin ml 5,000 8 Thuc nhum tm Gentian ml 5,000 9 Cn ty 95 ml 10,000 10 Lugol ml 5,000 11 Thuc nhum xanh methylen ml 5,000 12 Mi trng nui cy a 3,000 13 Panel nh danh theo h thng my t ng Test 1,000 14 B Panel chng B 0,0002 15 Bm kim tim Ci 1,000 16 Bng Kg 0,001 17 Cn 90 (v sinh dng c) ml 10,000 18 n cn Ci 0,0001 19 Panh Ci 0,0001 20 Khay ng bnh phm Ci 0,0001 21 Hp vn chuyn bnh phm Ci 0,0001 22 M Ci 0,020 23 Khu trang Ci 0,020 24 Gng tay i 3,000 25 Gng tay x l dng c i 0,020 26 Qun o bo h B 0,001 27 Acid ngm lam ml 10,000 28 ng nghim thy tinh ng 1,000 29 Bt vit knh Ci 0,020 30 Bt bi Ci 0,010 31 Bt la Ci 0,010 32 S lu kt qu xt nghim T 0,001 33 Cn st trng tay nhanh ml 1,000 34 Dung dch nc ra tay ml 8,000 35 Khn lau tay Ci 0,030 36 Giy tr kt qu xt nghim T 2,000 37 QC (nu thc hin) * 0,1 37 EQAS (nu thc hin) * 0,005 51. 36 B Panel s dng kim chun c tnh trn 5000 Test/1 nm. * Ghi ch: - Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng 10 mu cho 1 ln tin hnh k thut). - Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi s ln ngoi kim trung bnh 2 ln/1 nm). 3. Bnh phm Tt c cc loi bnh phm c ch nh nui cy tm vi khun gy bnh. 4. Phiu xt nghim in y thng tin theo mu phiu yu cu. III. CC BC TIN HNH Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn. 1. Ly bnh phm Theo ng quy nh ca chuyn ngnh Vi sinh (Xem ph lc 1). 2. Tin hnh k thut - Nhum soi bnh phm, nh gi s b - Nui cy bnh phm vo mi trng phn lp - m qua m - Bt khun lc nghi ng - Nhum soi, th nghim cc tnh cht sinh vt ha hc n gin v nh danh bng h thng t ng. IV. NHN NH KT QU - Dng tnh: Phn lp v nh danh c vi khun gy bnh. Tr kt qu tn vi khun n mc chi v/hoc loi. - m tnh: Khng tm thy hoc khng phn lp c vi khun gy bnh. V. NHNG SAI ST V X TR Quy trnh ny ch p dng tm vi khun hiu k kh ty tin d nui cy, khng p dng cho cc vi khun k kh bt buc. Kt qu m tnh khng c ngha l khng c vi khun gy bnh trong bnh phm m l khng tm thy cn nguyn vi khun gy bnh c th phn lp c bng quy trnh nui cy ny. Nu c yu cu tm cn nguyn vi khun no , phi ghi yu cu c th. Bnh phm ly, vn chuyn v bo qun khng ng yu cu c th a n kt qu m tnh hoc dng tnh gi (Xem Ph lc 6). 52. 37 12. Vi khun nh danh PCR I. MC CH V NGUYN L 1. Mc ch Chn on nhanh vi khun trc tip t bnh phm. 2. Nguyn l Xc nh s c mt ca gene c trng cho vi khun bng k thut PCR. II. CHUN B 1. Ngi thc hin - Ngi thc hin: Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh. - Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh. 2. Phng tin, ha cht Phng tin, ha cht nh v d di y hoc tng ng. 2.1. Trang thit b - T an ton Sinh hc cp 2 - My PCR - My in di - My c in di - My ly tm 25000 x g - My nhit - My vortex - My ly tm dng cho tube 0,2 ml - Micropipette - T m su (-200 C) - B lu in. 2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim) nh mc sinh phm v vt t tiu hao cho 2 mu/ln thc hin (VD) STT Chi ph ha cht, vt t tiu hao n v S lng 1 Bng kg 0,001 2 Cn ml 1,000 3 Bm kim tim Ci 1,000 53. 38 4 Panh Ci 0,0001 5 Khay ng bnh phm Ci 0,0001 6 Hp vn chuyn bnh phm Test 0,001 7 Tube ng bnh phm Ci 2,000 8 Gng khng c bt (DNase-RNase free) Ci 0,500 9 Sinh phm chn on Test 1,000 10 Khu hao sinh phm cho kim tra li Test 1,350 11 Kit tch chit DNA Test 2,350 12 DNA marker B 1,000 13 Primer 1 ml 0,0001 14 Primer 2 ml 0,0001 15 Ependoff 1,7ml Tube 3,000 16 Ependoff 0,2ml Tube 1,000 17 u cn 10 l c lc Ci 1,000 18 u cn 30 l Ci 1,200 19 u cn 200 l c lc Ci 2,200 20 u cn 1 ml c lc Ci 3,200 21 Ethanol BDH ml 0,500 22 Water-DEPC Treated ml 2,000 23 Giy thm Cun 0,100 24 Giy xt nghim T 2,000 25 S lu kt qu xt nghim T 0,001 26 Bt vit knh Ci 0,020 27 Bt bi Ci 0,010 28 M Ci 0,020 29 Khu trang Ci 0,020 30 Gng tay i 0,100 31 Gng tay x l dng c i 0,020 32 Qun o bo h B 0,005 33 Dung dch nc ra tay ml 8,000 34 Cn st trng tay nhanh ml 1,000 35 Dung dch kh trng ml 10,000 54. 39 36 Khn lau tay ci 0,010 37 QC (nu thc hin) * 0,100 38 EQAS (nu thc hin) * 0,005 * Ghi ch: - Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng 10 mu cho 1 ln tin hnh k thut). - Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi s ln ngoi kim trung bnh 2 ln/1 nm). 3. Bnh phm Vi khun thun v bnh phm phn lp 4. Phiu xt nghim in y thng tin theo mu phiu yu cu. III. CC BC TIN HNH Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn. 1. Ly bnh phm Theo ng quy nh ca chuyn ngnh Vi sinh (Xem ph lc 3). 2. Tin hnh k thut - Tch chit DNA tng s - Thc hin PCR - in di kim tra sn phm - nh gi v kt lun IV. NHN NH KT QU Sn phm PCR phi c mt bng c hiu duy nht, r nt, khng b t gy v c kch thc tng ng vi on gene ch cn khuch i. V. NHNG SAI ST V X TR Khi kt qu khi chng m v/hoc chng dng khng hp l, cn phi kim tra li qu trnh tch chit DNA tng s, cht lng primers v master mix, sau thc hin li xt nghim. 55. 40 13. Vi khun nh danh gii trnh t gene I. NGUYN L 1. Mc ch nh danh mi cn nguyn vi khun gy bnh n mc chi v hoc loi. 2. Nguyn l nh danh chnh xc cc loi vi khun da trn trnh t nucleotide c trng ca gene m ha cho 16S rRNA ca vi khun bng k thut gii trnh t gene. II. CHUN B 1.Ngi thc hin - Ngi thc hin: Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh. - Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh. 2. Phng tin, ha cht Phng tin, ha cht nh v d di y hoc tng ng. 2.1. Trang thit b - T an ton Sinh hc cp 2 - My PCR - My c in di - My gii trnh t gen - My ly tm 25000 x g - My nhit - My vortex - My ly tm dng cho tube 0,2 ml - Micropipette - B lu in 2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim) nh mc sinh phm v vt t tiu hao cho 3 mu/ln thc hin (VD) STT Chi ph ha cht, vt t tiu hao n v S lng 1 Bng kg 0,001 2 Cn ml 1,000