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Rapid H5-HA(Ab) ＲAPID TEST FOR DETECTION OF ANTIBODIES TO HEMAGGLUTININ (HA) OF INFLUENZA A VIRUS, H5 STRAIN
This kit is a chromatographic immunoassay for qualitative detection of antibodies to hemagglutining (HA) of Influenza Type-A viruses, H5 strain (also known as highly pathogenic avian influenza) in human body excretes, nasopharyngeal aspirates, chicken embryo whole virus inoculation or viral lysates, etc. It is intended for screening of for specific H5 HA antibodies and clinical identification of H5 avian influenza. The use of this chromatographic immunoassay provides rapid reliable and safe diagnostic method for detection of H5 avian influenza within 30 minutes. Simple, single use device that has integrated quality control band and do not require any additional laboratory equipment.
Avian influenza is an infectious disease of birds caused by type A strains of the influenza virus. The disease, first identified in Italy more than 100 years ago, now occurs worldwide. Infection triggers a wide spectrum of symptoms in birds, ranging from mild illness to a highly contagious and rapidly fatal disease resulting in severe epidemics. In the H5N1 bird flu in Hong Kong in 1997, patients had developed symptoms of fever, sore throat, cough and, in several of the fatal cases, severe respiratory distress secondary to viral pneumonia. Previously healthy adults and children, and some with chronic medical conditions, were affected. More recently, outbreaks of avian influenza H5N1 occurred among poultry in eight countries in Asia (Cambodia, China, Indonesia, Japan, Laos, South Korea, Thailand, and Vietnam) during late 2003 and early 2004. At that time, more than 100 million birds in the affected countries either died from the disease or were killed in order to try to control the outbreak. By March 2004, the outbreak was reported to be under control. Beginning in late June 2004, however, new deadly outbreaks of influenza H5N1 among poultry were reported by several countries in Asia (Cambodia, China, Indonesia, Malaysia [first-time reports], Thailand, and Vietnam ). It is believed that these outbreaks are ongoing. Human infections of avian influenza H5N1 however, have been reported in Cambodia (1case/1 death) Thailand (17cases/1 deaths) and Vietnam (51cases/ 33deaths) during both of these outbreak periods.
Hemagglutinin (HA) is a surface glycoprotein on Influenza A responsible for binding to N-AcetylNeuraminic Acid (NeuNAc) or commonly Sialic acid on host cell surface receptors. The Influenza viruses form the A virus group have principally similar morphological, chemical and biological features. The differentiation of the types is possible by the different antigenicity of their nucleo- and matrix proteins that have type-specific antigenicity. However, the essential immunodominant antigens and primary targets in diagnosis are the hemagglutinin (HA) and the neuraminidase (NA) antigens. Screening for type-specific anti-HA or anti-NA antibodies has also been proved to be useful method in clinical identification of different influenza strains.
H5-HA(Ab) Rapid Test employs chromatographic lateral flow device with antibody competitive –like principle for detection of hemagglutinin antibodies (anti-HA) in sample. Colloidal gold particles coated with complex of monoclonal antibodies reactive to H5 hemagglutinin (HA) and H5 lysate containing HA, are dry-immobilized onto a nitrocellulose membrane strip. When the sample is added, it migrates by capillary diffusion trough the strip rehydrating the gold conjugate. If present, anti-HA will bind with the HA coated onto the gold particles. The gold particles will continue to migrate along the strip until the Control Zone (C) where they are captured by goat, anti-mouse IgG antibodies previously immobilized there and a visible red line appears. If there is no H5 specific anti-HA in sample, the gold particles will be captured from monoclonal anti-HA antibodies previously immobilized on the Test Zone (T) and will aggregate as a red line .
Rapid H5-HA(Ab) card in aluminium pouch with desiccant: Package Insert -1 copy l Materials required but not provided: clock or timer, disposable pipettes, specimen collection container, centrifuge, biohazard waste container
1. Sample Collection: Either fresh serum or plasma samples can be used for this assay. Blood collected by venipuncture should be allowed to clot naturally and completely. Care should be taken to ensure that the serum samples are clear and not contaminated by microorganisms. Any visible particulate matters in the sample should be removed by centrifugation at 3000 RPM (round per minutes) for 20 minutes at room temperature or by filtration on 0.22u filters. Plasma samples collected into EDTA, sodium citrate or heparin may be tested, but highly lipaemic, icteric, or hemolized samples should not be used as they can give false results in the assay. Do not heat inactivate samples. This can cause sample deterioraration. 2. Transportation and Storage: Store samples at 2-8℃. Samples not required for assaying within 3 days should be stored frozen (-20℃ or lower). Avoid multiple freeze-thaw cycles.
This test can be stored at room temperature (18-30℃, do not freeze!) for 18 months from the date of manufacture (see label on strip pouch). Use immediately after opening.
This test is for In Vitro Use only FOR PROFESSIONAL USE ONLY 1. All the waste and sample should be treated in case of transmitting disease and must be properly disinfected (autoclaving is preferred) before disposal. 2. Once taking the strip out of the pouch, carry out your testing as early as possible (no more than 20minutes) to avoid moisture. The nitrocellulose membrane can absorb water, which can affect the test chromatography performance. 3. The performance characteristics of the test depend on sample quality and preparations. For strong positive samples, the red line corresponding to the Control Zone (C) may appear in 3-5 minutes after sample loading, but for weak positive samples, the red line may appear in the Control Zone(C) in 15 minutes. Results obtained after 30 minutes can lead to incorrect interpretation. 4. Make sure that the test is within the validity indicated. 5. Calibrate the pipette frequently to assure the accuracy. Use different disposal pipette tips for each specimen in order to avoid cross-contaminations. 6. Do not modify the test procedure. Avoid moisture. 7. A test giving an invalid result should be repeated. 8. Do not reuse lancets, test strips, pipettes. Autoclave before disposal. 9. All specimens from human origin should be considered as potentially infectious. Strict adherence to GLP (Good Laboratory Practice) regulations can ensure the personal safety. Never eat, drink, smoke, or apply cosmetics in the assay laboratory.
Allow the strip to reach room temperature (appropriately 30min). Open the pouch and slowly pipette 70μl of serum/plasma sample into the sample window (S). Avoid dropping solutions in the observation window. Place the strip on flat surface and read the results within 30minutes.
Positive Results: One red line appears in the Control Zone(C), indicating that antibodies to hemagglutinin of Influenza A viruses, H5 strain have been detected using this Rapid Test. Negative Results: Two red lines appear: one in the Control Zone (C), and additional line in the Test Zone (T) indicating that no antibodies to hemagglutinin of Influenza A, viruses H5 strain have been detected with this Rapid Test. However, this does not exclude the possibility for infection with H5 influenza virus. Quality Control: One red line always appears next to Control Zone (C). If no red line appears in the Control Zone (C), the test is invalid - discard the test and repeat with new sample and new strip
The positive result obtained with this H5-HA(Ab) Rapid Test alone cannot be the final diagnosis of infection with H5 strain avian influenza. Any positive result must be interpreted in conjunction with another laboratory testing results. Follow-up and supplementary testing with other analytical system is required to confirm any positive results.
REFERENCES: 1. Fouchier RAM, Schneeberger PM, Rozendaal FW, et al. Avian influenza A virus (H7N7) associated with human conjunctivitis and a fatal case of acute respiratory distress syndrome. Proc Natl Acad Sci 2004. Published online before print January 26, 2004 2. Fouchier RAM, Munster V, Wallensten A, et al. Characterization of a novel influenza A virus hemagglutinin subtype (H16) obtained from black-headed gulls J Virol 2005 Mar;79(5):2814-22 3. Horimoto T, Kawaoka Y. Pandemic threat posed by avian influenza A viruses. Clin Microbiol Rev 2001;14(1):129-49 4. Keawcharoen J, Oraveerakul K, Kuiken T, et al. Avian influenza H5N1 in tigers and leopards. Emerg Infect Dis 2004 Dec;10(12) 5. Luschow D, Werner O, Mettenleiter TC, et al. Protections of chickens from lethal avian influenza A virus infections by live-virus hemagglutinin (H5) gene. Vaccine 2001 Jul 20;19(30):4249-59 6. Monto AS. The threat of an avian influenza pandemic. (Perspective) N Engl J Med 2005 Jan 27;352(4):323-4 7. OIE. Highly pathogenic avian influenza. Technical disease card database. Apr 22, 2002
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