This kit is an enzyme-linked immunosorbent assay (ELISA) for qualitative detection of hemagglutinin (HA) of Influenza A Virus, H5 strain (also known as highly pathogenic avian influenza) in avian excretes, human nasopharyngeal aspirates,swabs, nasal wash, chicken embryo whole virus inoculation or viral lysates, etc. It is intended for clinical identification of specific H5, type-A influenza viruses. It is intended for clinical identification of the H5 strain of influenza A viruses.
PRINCIPLE OF THE ASSAY
H5-HA(Ag) ELISA kit uses polystyrene microwell strips pre-coated with antibodies (anti-HA"Ab") specific to the hemagglutinin (HA) of the H5 Influenza strain. Sample is added into the microwells. During the first incubation step, the specific anti-HA"Ab"-HA immunocomplex which forms in case of presence of H5 HA in the sample, is captured on the solid phase. After washing to remove sample proteins or unbound material, second H5 HA specific anti-HA"Ab" conjugated to horseradish peroxidase (anti-HA"Ab"HRP) is added. During the second incubations step, this anti-HA"Ab"HRP antibody will specifically bind to the anti-HA"Ab"-HA complex previously formed inside the wells.
Unbound anti-HA"Ab"HRP is removed after washing. Chromogen solutions containing tetramethylbenzidine (TMB) and urea peroxide are added into the wells.
In presence of the anti-HA "sandwich" immunocomplex, the colorless Chromogens are hydrolyzed by the bound HRP-conjugate to a blue colored product. The blue color turns yellow after stopping the reaction with sulfuric acid. The amount of color can be measured and is proportional to the amount of HA in the sample. Wells containing samples negative for H5 HA remain colorless."Ab" - HA - anti-HA"Ab"HRP
SENSITIVITY: primary study with cultured viruses
Detection of 14 different H5 viral strains collected from birds from affected areas in different province and regions in China (Including Hong Kong)
** HAU - One hemagglutination unit defined as the smallest virus concentration still causing a visible agglutination
ELISA detection limit : in 93% viral detection titre, sensitivity 0.25HA Units
Sensitivity and Specificity on different geographic located avian samples
Detect H5 (44x) strains collected worldwide
Negative results with H3 (2x), H7(2x), H9 (5x) Strains and Newcastle Diseases (2x) strains
Comparison of Sensitivity of H5 Antigen ELISA and Standard RT-PCR
SPECIFICITY : Large scalefield testing on poultry samples
400 previously tested as negative avian anal samples.
100 previously tested as negative avian oral samples.
Results: 1 false positive
PRODUCTION
ISO 9001:2000, ISO 13485:2003, GMP certificated
REFERENCES:
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
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
Horimoto T, Kawaoka Y. Pandemic threat posed by avian influenza A viruses. Clin Microbiol Rev 2001;14(1):129-49
Keawcharoen J, Oraveerakul K, Kuiken T, et al. Avian influenza H5N1 in tigers and leopards. Emerg Infect Dis 2004 Dec;10(12)
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 Monto AS. The threat of an avian influenza pandemic. (Perspective) N Engl J Med 2005 Jan 27;352(4):323-4
OIE. Highly pathogenic avian influenza. International Health Code. Chap 2.7.12. 2004
OIE. Update on avian influenza in animals in Asia (type H5). Updated frequently
Subbarao K, Klimov A, Katz J, Regnery H, Lim W, Hall H, et al. Characterization of an avian influenza A (H5N1) virus isolated from a child with a fatal respiratory illness. Science 1998;279:393–6.
Yuen KY, Chan PKS, Peiris M, Tsang DNC, Que TL, Shortridge KF, et al. Clinical features and rapid viral diagnosis of human disease associated with avian influenza A H5N1 virus. Lancet 1998;351:467–71.
For ordering, please go to DR.WANG, Fax: 0049 7071 792022, E-Mail: info@normae.de