QUALITY SYSTEM CERTIFIED ISO 9001 ISO 13485
LINEAR CHEMICALS S.L. Joaquim Costa 18 2ª planta. 08390 Montgat, Barcelona, SPAIN Telf. (+34) 934 694 990 Fax. (+34) 934 693 435. website www.linear.es
PRINCIPLE The LINEAR HA V IgM Rapid Te st i s a lateral fl ow chromatographi c immunoassay for the qualita tive dete ction of IgM antib odies to Hepati tis A virus (HAV) in hu man serum or pla sma. It i s in tended to be u sed a s a screening test and as an aid in th e diagnosis o f in fection w ith HAV. Any reactive specimen with the HAV ca ssete must be confirmed with alternative testing method(s) and clinical findings. The te st cassette con sists o f: 1) a burgundy color ed conj ugate pad containing mouse anti-human IgM anti body conjugated with colloidal gold (IgM conjugates) a nd 2) a nitrocellulose membrane strip con taining a test band (T band) and a control band (C band). The T band is pre-coa ted with recombinant HAV antigen, and the C band is pre- coated w ith goa t an ti-mouse IgM antibodies. When an a dequate v olume of test specimen i s disp ensed into the sample well of the cassette, the specimen m igrates by capillary action a cross the cassette. An ti-HAV IgM if p resent in the specimen w ill bind to the IgM conjugates. The im munocomplex is then captured on the membrane b y the pre-coated HAV antigen forming a burgundy colored T line, indicating a HAV IgM positive test result. Absence of the T line suggests a negative result. The test contains an internal control (C line) which should exhibit a burgundy colored line o f the immunocomple x of goat anti-m ouse Ig G/IgM-gold conjugate regardless of the col or development o f the T l ine. Otherwise, the test result is invalid and the specimen must be retested with another device.
PACKAGING CONTENTS
REF 4211240 40 HAV test device 40 Plastic dropper for HAV IgM 1 Sample diluent (5 mL)
STORAGE AND STABILITY Store at 2-3 0ºC. The test dev ice is stable through the expiration date
printed on the sealed pou ch. The te st device must re main in th e seale d pouch until use. Do not freeze the kit or expose the kit over 30°C. Do not use beyond the expiration date.
SPECIMEN COLLECTION AND PREPARATION
Plasma 1. Collect blood specimen into a lavender, blue or green top collection tube
(containing EDTA, citrate or heparin, respectively, in Vacuta iner® ) by veinpuncture.
2. Separate the plasma by centrifugation. 3. Carefully withdraw the plasma into new pre-labeled tube.
Serum 1. Collect blood specimen into a re d top collection tu be (containing no
anticoagulants in Vacutainer®) by veinpuncture. 2. Allow the blood to clot. 3. Separate the serum by centrifugation. 4. Carefully withdraw the serum into a new pre-labeled tube. Test specimens a s soon as po ssible after colle cting. Store at 2-8°C if not tested immediately, for up to 5 day s. The specimens should be frozen a t -20°C for longer storage. Avoid multiple freeze-thaw cycles. Prior to testing, bring fro zen specimens to room temperature slowly and mix gently . Specimens containing v isible parti culate ma tter should be clar ified by centrifugation. Do not use samples demonstr ating gr oss lip emia, gro ss hemolysis or turbidity in order to avoid interference on result interpretation.
MATERIAL REQUIRED
- Timer
PROCEDURE
Allow the test cassette, urine or serum specimen and/or controls to equilibrate to room temperature (20-30°C) prior to testing.
1. Remove the test from its sealed pou ch, and place i t on a clean , lev el surface. Label the device w ith patien t or control iden tification. For be st results the assay should be performed within one hour.
2. Fill the blood transfer device with the specimen (serum, plasma or whole blood).
3. Holding the p lastic dropper vertically, dispense 1 d rop specimen (30-45 l serum) into the sample well (S well), making sure that there are no air bubbles. Then hold the diluent bottle vertically and add 1 drop (about 35-50 µL) of sample diluent immediately.
4. Set up timer. 5. Results can be r ead after 15 minutes. Positive results can be v isible in
as soon as 1 minute.
Do not read results after 15 minutes. To avoid confusion, discard the test device after interpreting the result.
INTERPRETATION OF ASSAY RESULT
NEGATIVE RESULT
If only the C line is dev eloped, the test indicates that no detectable IgM anti-HAV is present in the specimen. The result is negative.
POSITIVE RESULT
If both the C and the T lines are developed, the test indicates the presence of IgM anti-HAV in the specimen. The result is positive.
Samples with positive results should be confirmed with alternative testing method(s) and clinical findings before a positive determination is made.
INVALID
If no C line is developed, the assay is invalid regardless of color development on the T line as indicated below. Repeat the assay with a new device.
CONTENTS
For professional in vitro diagnostic use only
REF
HAV IgM A rapid one test for the qualitative detection of IgM antibodies to Hepatitis A virus (HAV) in human serum or plasma. ONE STEP
4211240 HAV 40 Tests
HAV cassette
QUALITY SYSTEM CERTIFIED ISO 9001 ISO 13485
LINEAR CHEMICALS S.L. Joaquim Costa 18 2ª planta. 08390 Montgat, Barcelona, SPAIN Telf. (+34) 934 694 990 Fax. (+34) 934 693 435. website www.linear.es
QUALITY CONTROL
A procedural control is included in th e test. A colored line appearing in the control line region (C) is considered an internal procedural control. It confirms sufficient sp ecimen v olume, adequate membrane w icking and correct procedural te chnique. Control standards are no t sup plied w ith this ki t; however, it i s recommended that positive and negative controls be tested as good labora tory te sting pra ctice to confirm the test p rocedure and to verify proper test performance.
CLINICAL SIGNIFICANCE
HAV is a p ositive-sense RNA virus, a unique member of picornaviridae1. Its transmission depends primarily on serial transmission from person to per son by the fecal-oral r oute. Altho ugh he patitis A i s not or dinarily a sex ually transmitted disease, the infection ra te is high amo ng male homosexuals as result of oral-anal contact2,3.
The presence of specific anti-HAV IgM in blood sample s suggests acute or recent HAV in fection4-6. The IgM a ntibody rapidly incre ases in titer over a period of 4-6 weeks post infection and then declines to non-detectable levels within 3 to 6 months in most patients7.
The HAV casse tte is to be used to det ect IgM an ti-HAV in less than 15 minutes by untrained or minimally skilled person nel, w ithout cumbe rsome laboratory equipment.
ANALYTICAL PERFORMANCE
A total o f 200 samples from su sceptible subje cts w ere tested by the HAV cassette and by a commercia l EIA test. Compari son o f the re sults for a ll subjects is shown in the following table:
HAV IgM Rapid Test EIA Positive Negative Total
Positive 21 1 22 Negative 0 178 178
Total 21 179 200 Relative Sensitivity: 95.5% , Relative Specificity: 100%, Overall Agreement: 99.5%
LIMITATIONS OF TEST 1. The Assay Procedure and the In terpretation o f Assay Resul t sections
must be followed closely when testing for the presence of anti-HAV IgM in ser um or pla sma from ind ividual subje cts. Failu re to follow the procedure may give inaccurate results.
2. The HAV cassette is limited to the qualitative detection of anti-HAV IgM in human serum or plasma. The in tensity of the test line does not have linear correlation with the antibody titer in the specimen.
3. A nega tive resul t for an i ndividual subj ect in dicates abse nce o f detectable anti-HA V IgM. How ever, a negativ e test re sult do es no t preclude the possibility of exposure to or infection with HAV.
4. A negative result can occur if the quantity of the anti-HAV IgM present in the spe cimen is below the detec tion limits of the assay or the antibodies that are detected are not present during the stage of disease in which a sample is collected.
5. Some sp ecimens con taining unu sually high ti ters of he terophile antibodies or rheumatoid factor may affect expected results.
6. The resul ts obtain ed w ith thi s te st should only be interpre ted i n conjunction with other diagnostic procedures and clinical findings.
PRECAUTIONS 1. This package insert must be read com pletely before performing
the test. Failure to follow the insert gives inaccurate test results.2. Do not open the sealed p ouch unle ss ready to con duct the
assay. 3. Do not use expired devices. 4. Bring all reagen ts to room temperature (15°C - 30°C) before
use. 5. Do not use compo nents from any other ty pe of te st kit as a
substitute for the components in this kit. 6. Do not use hemolized blood specimens for testing. 7. Wear protective clothing and di sposable gloves while handling
the kit reagents and clinical specimens. Wash hands thoroughly after performing the test.
8. Users o f this test should f ollow t he US CDC Uni versal Precautions for pr evention of transmission o f HIV , HB V and other blood-borne pathogens.
9. Do not smo ke, dri nk or ea t in are as w here specimen s or ki t reagents are being handled.
10. Dispose of all specimens and materials used to perform the test as bio-hazardous waste.
11. Handle the n egative and po sitive controls in the same manner as patient specimens.
12. The te st re sults should be read w ithin 15 minu tes a fter a specimen is a pplied to the sample w ell or sample pa d of th e device. Reading the resu lts after 15 minutes ma y gi ve erroneous results.
13. Do not perform the test in a r oom with strong air flow, i.e. an electric fan or strong air-conditioning.
REFERENCES
1. Minor P. Picornaviridae. In: Francki RIB, Fauquet CM, Knudson DL, et al., eds. Classification and nomenclature of viruses (Arch Virol Supp 2). Wien: Springer-Verlag, 1991: 320-326.
2. Keeffe EB . Clini cal approach to v iral hepatitis in homo sexual men. Med Clin North Am. 1986;70(3):567-86.
3. Ballesteros J, Dal-Re R, Gonzalez A, del Romero J. Are homosexual males a ri sk group for hepatiti s A in fection in intermediate end emicity areas? Epidemiol In fect. 1996 ; 117(1):145-8.
4. Bradley DW, Maynard JE, Hindman SH, et al: Serodiagnosis of viral hepatiti s A : D etection of a cute-phase immunogl obulin M anti-hepatitis A v irus by radioimmunoassay . J Clin Microbiol 1977; 5: 521-530.
5. Decker RH, Kosakowski SM, Vanderb ilt AS, et al: Diagnosis of acute hepatitis A by HAVAB-M : A dir ect radioimmunoassay for IgM anti-HAV. Am J Clin Pathol 1981;76:140-147.
6. Locarnini SA, Fer ris AA , Lehman NI, et al : The an tibody response following hepati tis A i nfection. In tervirology 1974; 4:110-118.
7. Skinhoj P, Mikkelsen F, Holli nger FB. Hepatitis A in Greenland : Importance o f sp ecific an tibody testin g in epidemiologi c surveillance. Am J. Epidemiol 1977; 105: 104-147.
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