summary of the diagnostic value of specific microbiological investigations
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Summary of the diagnostic value of specific microbiological investigations. Blood culture. Technique Minimal 1~3 mL each Different venipuncture ( ≥ 2 sets) Central catheter + Peripheral culture Colonized bacteria – possibility of contamination. Blood culture. Blood culture. Gram stain. - PowerPoint PPT PresentationTRANSCRIPT
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Summary of the diagnostic value of specific microbiological investi-
gations
Test Diagnostic value
False posi-tive rate
False nega-tive rate
Blood culture ++++ - +++ Viral antigen detection (nasopharyngeal aspirate) +++ - +
Viral culture +++ - ++ Serum antigen ++ + ++ Urine antigen + ++ ++ Paired antibody titre +++ + ++ Bacterial culture of na-sopharyngeal secretions
- +++ +
Lung puncture culture ++++ - +
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Blood culture• Technique
– Minimal 1~3 mL each– Different venipuncture (≥2 sets)– Central catheter + Peripheral culture
• Colonized bacteria – possibility of contami-nation
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Blood culture
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Blood culture
Gram stain
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Interpretation of blood culture• Almost always
important– S. aureus– S. pneumoniae– S. pyogenes (GAS)– S. agalactiae (GBS)– H. influenzae– Enterobacteriaceae
(E. coli, Klebsiella, etc..)– Bacteroidaceae– P. aeruginosa– Candida species
• Possible contaminant– P. acnes– Corynebacterium species– Bacillus species– CoNS (coagulase negative
Staphylococci, S. epider-midis)
– S. viridans– Peptostreptococcus
Most causative pathogens are compatible with clinical syn-drome
Most causative pathogens will grow within 72 hours
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Nasopharyngeal aspirate collection
• Nasopharyngeal aspirate – Virus culture / PCR
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Early diagnosis of viral infection
Microscopy – LM– EM
Detecting antigen– IF staining– Solid phase immunoassay (ELISA, RIA, LA), – Immunochromatography
Detecting nucleic acid– nucleic acid amplification (PCR, etc.), – nucleic acid hybridization
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Tzank test of HSV Vesicular Lesion
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Ultrastructural Characteristics of SARS-Asso-ciated Coronavirus Grown in Vero E6 Cell
100 nm
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Indirect Immunofluorescent staining
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Immunofluorescent staining
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• Indirect IF Staining for RSV, • Specimen: Nasal aspirate
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• Indirect IF Staining for PIV3 • Specimen: Nasal aspirate• Confocal microscopy
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Antigen Detec-tion
by ELISA
Direct Method
Indirect Method
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Indirect ELISA for RSVSpecimen: Nasal aspi-rate
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Immunochromatography
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Conventional method vs. RT-PCR
Virus identified No. detected by RT-PCR(% of total episodes)
Respiratory syncytial virus 122 (23.7) 596 (14.7)Adenovirus 35 (6.8) 243 (6.0)Parainfluenza virus type1 11 (2.1) 30 (0.7)
type 2 ND 23 (0.6)
type 3 32 (6.2) 195 (4.8)
type 4 ND 2 (0.05)Influenza virus A 24 (4.7) 117 (2.9)
B 9 (1.8) 47 (1.2)
Total 316 (61.4) *
*More than 1 virus were isolated from 36 patients (11.4%) among 514 patients, Sep 2000-Aug 2005.**1,270 strains of viruses were isolated from 1,234 out of 4,058 cases (30.4% of total patients), Nov 1990-May 2002. More than 1 virus were isolated from 36 patients.
Detection by conventional method (% of total episodes)
1,270 (30.4) **
RhinovirusMetapneumovirusCoronavirus, conventionalCoronavirus NL63BocavirusUnknown
40 (7.8)26 (5.0) 1 (<1) 9 (1.8)58 (11.2)
ND ND ND ND ND 17 (0.3)
Choi EH et al. Clin Infect Dis 2006;43:585-92