Evaluation of rapid detection kit against avian influenza A virus and H5 subtype for field Sample


Michael Haryadi Wibowo(1*), Tri Untari(2), Sidna Artanto(3), Krisdiana Putri(4), Surya Amanu(5), Widya Asmara(6)

(1) Department of Microbiology, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia
(2) Department of Microbiology, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia
(3) Department of Microbiology, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia
(4) Department of Public Health, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia
(5) Department of Microbiology, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia
(6) Department of Microbiology, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia
(*) Corresponding Author


Avian influenza virus is poultry viral disease, which causes high economic losses. Various efforts have been made to control the disease. One effort is required screening fast and precise diagnostic test. This study was aimed to determine the potential of rapid test kit of AIV/H5 Anigen Rapid Test for the detection of AI virus types A and subtype H5 in the field. Some tests were carried out, e.g.: the potential test, cross-reaction test, sensitivity and specificity test. Potency test was done to evaluate potential of detection limits of the kit, by having the test of serial dilution of AI virus positive control. Cross-reaction test was done to detect antigens other than AI virus H5N1, e.g.:  IB virus of Massachuset strain, IBV strain 4-91, Newcastle Disease virus, and Escherichia coli. Sensitivity and specificity test were applied to the filed samples which clinically and laboratory were confirmed as H5N1 positive. To confirm the result of rapid test was then being done by reverse transcriptase polymerase chain reaction. Based on these results it can be concluded that, Anigen Kit AIV/H5 Ag Rapid Test can detect antigen-containing samples having AI virus HA titer up to 26of type A virus, and up to 25 for subtype H5 virus. Anigen Kit AIV/H5 Ag Rapid Test showed no cross-reactions with Infectious Bronchitis virus, Newcastle Disease virus, and Escherichia coli. Anigen A Rapid Test Kit AIV Ag showed a sensitivity of 50% and specificity of 100%, while Anigen Ag Rapid Test Kit AIV/H5 showed a sensitivity of 25% and specificity is 100%.


avian influenza; rapid detection kit; specificity and sensitivity

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Anonymous, 2003. Avian Influenza. Preliminary Final Report/OIE Terrestrial Animal Health Standards Commision. Appendix. Pp. 7-11.

Asmara W., Wibowo M.H., Tabbu C.R., 2005. Hemagglutini Subtype Identification of Avian Influenza Virus Isolated from Various Species of Birds Using RT-PCR. Sain Vet.J. : 42-45.

Charlton B., Crossley B., and Hietala S. 2009. Conventional and Future Diagnostics for Avian Influenza. Comp. Immunol. Microbiol. and Infect. 32:341-350.

Chen H., Smith G.J.D., Li K.S., Wang J., Fan X. H., Rayner M. J., Vijaykrishna D., Zhang J.X., Zhang L.J., Guo C. T., Cheung C. L., Xu K.M., Duan L., Huang K., Qin K., Leung Y.H.C., Wu W.L., Lu H.R., Chen Y., Xia N.S., Naipospos T.S.P.,Yuen K.Y., Hassan S.S., Bahri S., Nguyen T.D., Webster R.G., Peiris J.S.M., and Guan Y. 2006. Establishment of Multiple Sub lineages of H5N1 Influenza Virus in Asia: Implication for Pandemic Control. PNAS 103 (8): 2845-2850.

Chua T-H., Ellis, T.M., Wong C.W., Guan Y., Ge, S.X., Peng G., Lamichane C., Maliadis C., Tan S-W., Selleck P., and Parkinson J., 2007. Performance Evaluation of Five Detection Test for Avian Influenza Antigen with Various Avian Sample. Avian Dis. 56: 96-105.

Cox N.J. and Kawaoka Y. 1998. Orthomyxoviruses: Influenza. Dalam: Microbiology and microbial infection. Ed Collier, L., Ballows, A. And Sussman, M. Vol.1: Virology, Oxford University press, Inc.New York. Hal. 386-433.

Damayanti R., Dharmayanti, N.L.P.I., Indriani R., Wiyono A., and Darminto, 2004. The clinico-pathological Effect of Chicken Infected with Highly Pathogenic Avian Influenza in Some Farms in East Java and West Java. JITV 9(2): 128-135.

Dharmayanti N.L.P.I, Damayanti R., Wiyono A., Indriyani R., and Darminto. 2004. Identification of Avian Influenza Virus of Indonesian Isolate by Reverse Transcriptase Polymerase Chain Reaction. JITV 9(2): 136-143.

Dharmayanti N.L.P.I., and Indriyani R., 2007. Molecular Pathogenicity of Avian Influenza Virus Isolated in the Year 2005. Media Kedokteran Hewan 23(2): 68-73.

Fouchier R.A.M., Munster V., Waltensen A., Besterbroer T.M., Herfst S.., Swith D., Rimnelzwaan G.F., Olsen B. And Osterhaus A.D.M.E. 2005. Characterization of Novel Influenza A Virus Hemagglutinin subtype (H16) Obtain from Black Headed Gulls. J. Virol 79 (5): 2814-2822.

Gambaryan A., Tuzikov A., Pazynina G., Bovine N., Balish A., Klimov A. 2005. Evolution of the receptor binding phenotype of influenza A (H5) viruses. Virology. 344: 432:438.

Hidayanto N.K., Asmara W., and Wibowo M.H., 2015. Characterization of Avian Influenza Virus H5N1 Subtype of Non Structural Gene from Duck Isolated in 2013. Sain Vet.J. 33(2): 195-204.

Ito I., Goto H., Yamamoto E., Tanaka H., Takeuchi M., Kuwayama M., Kawaoka Y., and Otsuki K. 2001. Generation of HPAI A Virus from an Avirulent Field Isolate by Passaging in Chicken. J. Virol. 75. (9): 4439-4443.

Kang B., Oh J., Lee C., Park B., Park Y., Hong K., Lee K., Cho B., and Song D. 2007. Evaluation of a Rapid Immunodiagnostic Test Kit for Rabies Virus. J. Vir. Method. 145: 30-36.

Khewcharoen J., Oraveerakul K., Kuiken T., Fourchier R.A., Amonsin A., Payungporn S. 2004. An Avian Influenza H5N1 in Tiger and Leopard. Emerg. Infect.Dis. 10: 2189-2191.

Loth L., Prijono W.B., Wibawa H., dan Usman T.B., 2008. Evaluation of Two Avian Influenza Type A Rapid Antigen Tests Under Indonesian Field Conditions. Diagn. Invest. 20: 642-644.

Mahardika G.N., Jonas M., Murwijati T., Fitria N., Suartha I.N., Suartini I.G.A.A., Wibawan I.W.T. 2016. Molecular Analysis of Hemmagglutinin-1 Fragment of Avian Influenza H5N1 Viruses Isolated from Chicken Farms in Indonesia from 2008 to 2010. Vet. Microbiol. 186: 52-58.

Martin S.W., Meek A.H., and Willeberg P. 1988. Sensitivity and Specificity. In Veterinary Epidemiology Principles and Methods. Iowa State University Press. p.:63-66.

National Committee for Avian Influenza and Preparedness of Influenza Pandemic, (NCAIPIP), 2008. The Timeline Data of Avian Influenza Outbreak in Indonesia. http//www.komnasfbpi.go.id/situation/update.html.

Payungporrn S., Piraya P., Salin C., Apiradee T., Juthatip K., Kanisak O.V., Alongkorn A., and Yong P. 2004. Single-Step Multiplex Reverse Transcriptase-Polymerase Chain Reaction for Influenza A Virus Subtype H5N1 Detection. Viral Imm. 17: 588-593.

Peng F., Wang Z., Zhang S.,Wu R., Hu S., Li Z., Wang X., dan Bi D. 2008. Development of An Immunochromatographic Strip For Rapid Detection Of H9 Subtype Avian Influenza Viruses. J. Clinical and Vaccine Immunology, 15 (3): 569–574.

Smith G.J.D., Naipospos T.S.P., Nguyen T.D., de Jong M.D., Vijaykrishna D., Usman T.B., Hassan S.S., Nguyen T.V., Dao T.V., Bui N.A., Leung Y.H. C., Cheung C.L., Rayner J.M., Zhang J.X., Poon L.L. M., Li K.S., Nguyen V.C., Hien T.T., Farrar J., Webster R.G., Chen H., Peiris J.S.M., and Guan Y. 2006. Evolution and Adaptation of H5N1 Influenza Virus in Avian and Human Hosts in Indonesia and Vietnam. Virol. 350: 258-268.

Sellect P., Avian Influenza Rapid Test Field Trial Data. Australian Animal Health Laboratory, Geelong, Australia. Un-published data. Songserm T., Alongkorn A., Rungroj J., Namdee S.H., Noppodal M., Nuananong P., Sunchai, P. 2006. Avian Influenza H5N1 in Naturally infected Domestic Cat. Emerg. Infect. Dis.www.cdc.gov/eid. 12 (11): 1744-1764.

Srihanto E.A., Asmara W., and Wibowo M.H., 2015. Philogenetic Molecular and Antigenic Structure Analysis of Avian Influenza Virus of Subtype H5N1 Lampung Isolates Collected during 2008 to 2013. Jurnal Kedokteran Hewan 9(1): 83-88.

Suarez D.L., 2003. Moecular Diagnostic Techniques: Can We Identify Influenza Viruses, Differentiate Subtype and Determine Pathogenicity Potential of Viruses by RT-PCR. Avian Dis. 47: 318-325.

Susanti R., Soejoedono R.D., Mahardika I.G.N.K., Wibawan I.W.T.,Suhartono M.T. 2008. Molecular Analysis of Hemagglutinin Gene of HPAI of H5N1 Subtype Isolated from Waterfowl. JITV 13(3): 229-239.

Tantilertcharoen R., Prapasakul N., Thanawongnuwes R., and Kramomthong I., no year. Performance Evaluation of Avian Influenza Virus Antigen Rapid Test Kit. Veterinary Diagnostic Laboratory Unit, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330.

Tong S., Li Y., Rivailler P., Christina C., Danilo A., Castillo A., Chen L.M., Recuenco S., Ellison J.A., David C.T., York I.A., Turmelle A.S., Moran D., Rogers S., Shi M., Tao Y., Weil M.R., Tong K., Rowe L.A., Sammons S., Xu X., Frace M., Lindblade K.A., Cox N.J., Anderson L.J., Rupprecht C.E., and Donis O., 2012. A Distinct Lineage Influenza A Virus from Bats. Proc. Nat.Aca.Sci. 109: 4269-4274.

Tsuda H., Sakoda Y., Sakabe S., Mochizuki T., Namba Y., dan Kida H. 2007. Development of an Immunochromatographic Kit for Rapid Diagnosis of H5 Avian Influenza Virus. Infection.J. Microbiol. Immunol., 51(9) 2007.

Wibawa H., Prijono W.B., Dharmayanti N.L.P.I., Irianingsih S.H., Miswati Y., Rohmah A., Andesyha E., Romlah, Daulay R.S.D., and Safitria K. 2012. Outbreak investigation of Duck Disease in Central Java Province, Yogyakarta Special Province, and East Java Province: Identification of New Clade of Avian Influenza Virus H5N1 Subtype in Indonesia. Buletin Laboratorium Veteriner 12 (4):1-9.

Wibowo H.W., Susetya H., Untari T., Putri K., Tabbu C.R., and Asmara W. 2006. Molecular Study on the Pathogenicity of Avian Influenza Virus. Indo. J. of Biotech. 11(2): 901-907.

Wibowo M.H., Susetya H., Untari T., Wahyuni A.E.T.H., Tabbu C.R., and Asmara W., 2007. Molecular Identification of Avian Influenza Virus from Cases Both with and without Symptom Specific for Avian Influenza. J. Vet. 8(3): 103-110.

Wibowo M.H., Srihanto E.A., Putri K., Asmara W., and Tabbu C.R. 2013. The Development of Pathogenicity of Avian Influenza Virus Isolated from Indonesia. Indo.J.of Biotech. 18(2): 133-143.

Wibowo M.H., Anggoro D., Amanu S., Wahyuni A.E.T.H., Untari T., Artanto S., and Asmara W., 2017. Receptor Binding and Antigenic Site Analysis of Hemagglutinin Gene Fragment of Avian Influenza Virus Serotype H5N1 Isolated from Indonesia. Pak. Vet. J. 37(2): 123-128.

Yamamoto Y., Nakamura K., Okamatsu M., Miyazaki A., Yamada M., and Mase M. 2008. Detecting Avian Influenza Virus (H5N1) in Domestic Duck Feathers. Emerg. Infect. Dis. 14(10): 1671-1672.

Wiyono A., Indriani R., Dharmayanti N.L.P.I., Damayanti R., Parede L., Syafriati T., and Darminto, 2004. Isolation and Characterization of HPAI virus H5 Subtype of Chicken from Outbreak in Indonesia. JITV 9(1): 61-71.

DOI: https://doi.org/10.22146/ijbiotech.26792

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