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| Development of a cELISA for effective detection of the antibody against H7 subtype of avian influenza virus |
| WANG Cong-cong1, WANG Si-wen1, ZHANG Ying1, 2, SHI Jian-zhong1, YIN Xin1, LI Cheng-jun1, WANG Xiu-rong1 |
1 State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, P.R.China
2 Shenyang Institute of Technology, Shenyang 110870, P.R.China
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摘要
【目的】H7亚型禽流感病毒(Avian influenza virus, AIV)和H5亚型一样，多数属于高致病性毒株，通常在禽类间传播，引起家禽出现严重的临床症状和高死亡率。从1996年至2012年，加拿大、意大利、墨西哥、荷兰、英国和美国出现了人感染H7亚型流感病毒(H7N2、H7N3和H7N7)的病例。2013年，中国报告了人感染H7N9流感病毒的事件，此后，该病毒持续在人和禽类中传播。因此，开展H7亚型禽流感病毒抗体检测工作成为一个重要问题。【方法】该研究用纯化的H7-AIV作为包被抗原，辣根过氧化物酶（Horseradish peroxidase, HRP）标记的单克隆抗体1H9（HRP-1H9）作为竞争抗体，采用棋盘滴定法建立了一种检测H7亚型AIV抗体的竞争性酶联免疫吸附实验(Competitive enzyme-linked immunosorbent assay, cELISA)。【结果】抗原包被的最佳浓度为5 μg mL-1，血清稀释度为1/10，酶标抗体为1/3000。用ROC曲线分析法测定178份AIV阴性和368份AIV阳性血清（n=546）的cELISA临界值，当PI为40%时，cELISA的特异性和敏感性分别为99.15%和98.12%。该方法可检测H7Nx（N1--N4、N7--N9）禽流感病毒抗体，与H1 -- H6、H8 -- H15亚型禽流感病毒及鸡新城疫病毒(Newcastle disease virus, NDV)、传染性支气管炎病毒(Infectious bronchitis virus, IBV)和传染性法氏囊病病毒(Infectious bursal disease virus, IBDV)等常见禽类病毒无反应，具有良好的特异性。该方法与血凝素抑制试验的符合率为98.56%。重复性实验表明，批内和批间重复的变异系数均小于12%。【结论、创新性】综上所述，本研究利用H7亚型特异性酶标单克隆抗体作为竞争抗体，建立了cELISA抗体检测方法，其优势是酶标抗体与待检血清同时加入反应孔中进行竞争反应，与先加入待检血清，再加竞争抗体反应的常规方法比较，简化了操作步骤，缩短反应时间，为开展H7-AIV抗体的大量检测提供了一种简单、便捷的技术手段。
Abstract H7 avian influenza viruses (AIVs) normally circulated among birds before. From 1996 to 2012, human infections with H7 AIVs (H7N2, H7N3, and H7N7) were reported in Canada, Italy, Mexico, the Netherlands, the United Kingdom and the USA. Until March 2013, human infections with H7N9 AIVs were reported in China. Since then, H7N9 AIVs have continued to circulate in both humans and birds. Therefore, the detection of antibodies against the H7 subtype of AIVs has become an important topic. In this study, a competitive enzyme-linked immunosorbent assay (cELISA) method for the detection of antibody against H7 AIVs was established. The optimal concentration of antigen coating was 5 μg mL–1, serum dilution was 1/10, and enzyme-labeled antibody was 1/3 000. To determine the cut-off value of cELISA, percent inhibition (PI) was determined by using receiver operating characteristic (ROC) curve analysis in 178 AIVs negative samples and 368 AIVs positive serum samples (n=546). When PI was set at 40%, the specificity and sensitivity of cELISA were 99.4 and 98.9%, respectively. This method could detect the antibodies against H7Nx (N1–N4, N7–N9) AIVs, and showed no reaction with AIVs of H1–H6 and H8–H15 subtypes or common avian viruses such as Newcastle disease virus (NDV), Infectious bronchitis virus (IBV) and Infectious bursal disease virus (IBDV), exhibiting good specificity. This method showed a coincidence rate of 98.56% with hemagglutinin inhibition (HI) test. And the repeatability experiment revealed that the coefficients of variation (CV) of intra- and inter-batch repetition were all less than 12%. The data indicated that the cELISA antibody-detection method established in this study provided a simple and accurate technical support for the detection of a large number of antibody samples of H7-AIV.
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Received: 06 January 2020
Accepted: 01 February 2021
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| Fund: This study was supported by the National Key R&D Program of China (2016YFD0500800). |
| About author: WANG Cong-cong, Tel: +86-451-51051684, E-mail: 1123051396@qq.com; Correspondence WANG Xiu-rong, Tel: +86-451-51051680, Fax: +86-451-51997166, E-mail: wangxiurong@caas.cn |
Cite this article:
WANG Cong-cong, WANG Si-wen, ZHANG Ying, SHI Jian-zhong, YIN Xin, LI Cheng-jun, WANG Xiu-rong.
2022.
Development of a cELISA for effective detection of the antibody against H7 subtype of avian influenza virus. Journal of Integrative Agriculture, 21(1): 199-207.
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Ahn S J, Baek Y H, Lloren K K S, Choi W S, Jeong J H, Antigua K J C, Kwon H I, Park S J, Kim E H, Kim Y I, Si Y J, Hong S B, Shin K S, Chun S, Choi Y K, Song M S. 2019. Rapid and simple colorimetric detection of multiple influenza viruses infecting humans using a reverse transcriptional loop-mediated isothermal amplification (RT-LAMP) diagnostic platform. BMC Infectious Diseases, 19, 676.
Bisset A T, Hoyne G F. 2020. Evolution and adaptation of the avian H7N9 virus into the human host. Microorganisms, 8, 778.
Chen L, Ruan F, Sun Y, Chen H, Liu M, Zhou J, Qin K. 2019. Establishment of sandwich ELISA for detecting the H7 subtype influenza A virus. Journal of Medical Virology, 91, 1168–1171.
Dong J, Fan J, Wang Y, Zhang Q, Yang Y, Jia Y, Ming F, Zhang X, Yao R, Li S, Zhang L, Li R, Xu G. 2019. Development and evaluation of a C-ELISA for rapid detection of antibody to AIV-H7. Analytical Biochemistry, 572, 52–57.
Gerber M, Isel C, Moules V, Marquet R. 2014. Selective packaging of the influenza A genome and consequences for genetic reassortment. Trends in Microbiology, 22, 446–455.
He F, Kiener T K, Lim X F, Tan Y, Raj K V, Tang M, Chow V T, Chen Q, Kwang J. 2013a. Development of a sensitive and specific epitope-blocking ELISA for universal detection of antibodies to human enterovirus 71 strains. PLoS ONE, 8, e55517.
He F, Prabakaran M, Tan Y, Indira K, Kumar S R, Kwang J. 2013b. Development of dual-function ELISA for effective antigen and antibody detection against H7 avian influenza virus. BMC Microbiology, 13, 219.
Imai M, Watanabe T, Kiso M, Nakajima N, Yamayoshi S, Iwatsuki-Horimoto K, Hatta M, Yamada S, Ito M, Sakai-Tagawa Y, Shirakura M, Takashita E, Fujisaki S, McBride R, Thompson A J, Takahashi K, Maemura T, Mitake H, Chiba S, Zhong G, et al. 2017. A highly pathogenic avian H7N9 influenza virus isolated from A human is lethal in some ferrets infected via respiratory droplets. Cell Host & Microbe, 22, 615–626.
Inui K, Nguyen T, Tseng H J, Tsai C M, Tsai Y L, Chung S, Padungtod P, Zhu H, Guan Y, Kalpravidh W, Claes F. 2019. A field-deployable insulated isothermal RT-PCR assay for identification of influenza A (H7N9) shows good performance in the laboratory. Influenza and Other Respiratory Viruses, 13, 610–617.
Isakova-Sivak I, Rudenko L. 2017. Tackling a novel lethal virus: A focus on H7N9 vaccine development. Expert Review of Vaccines, 16, 1–13.
Jiang L, Zhao X N, Xu W, Zhou X H, Luo C R, Zhou J N, Fu X Q, Chen Y Y, Li D. 2020. Emergence of human avian influenza A(H7N9) virus infections in Wenshan City in Southwest China, 2017. BMC Infectious Diseases, 20, 154.
Karawita A C, Tong M Z W, Short K R. 2019. A delicate balancing act: Immunity and immunopathology in human H7N9 influenza virus infections. Current Opinion in Infectious Diseases, 32, 191–195.
Liu L, Zhang Y, Cui P F, Wang C C, Zeng X Y, Deng G H, Wang X R. 2019. Development of a duplex TaqMan real-time RT-PCR assay for simultaneous detection of newly emerged H5N6 influenza viruses. Virology Journal, 16, 119.
Liu L T, Wang T, Wang M Y, Tong Q, Sun Y P, Pu J, Sun H L, Liu J H. 2019. Recombinant turkey herpesvirus expressing H9 hemagglutinin providing protection against H9N2 avian influenza. Virology, 529, 7–15.
Mythili T, Rajendra L, Bhavesh T, Thiagarajan D, Srinivasan V A. 2011. Development and comparative evaluation of a competitive ELISA with rose bengal test and a commercial indirect ELISA for serological diagnosis of brucellosis. Indian Journal of Microbiology, 51, 528–530.
Nelson M I, Vincent A L. 2015. Reverse zoonosis of influenza to swine: New perspectives on the human–animal interface. Trends in Microbiology, 23, 142–153.
Perez Rubio A, Eiros J M. 2018. Cell culture-derived flu vaccine: Present and future. Human Vaccines & Immunotherapeutics, 14, 1874–1882.
Prabakaran M, Ho H T, Prabhu N, Velumani S, Szyporta M, He F, Chan K P, Chen L M, Matsuoka Y, Donis R O, Kwang J. 2009. Development of epitope-blocking ELISA for universal detection of antibodies to human H5N1 influenza viruses. PLoS ONE, 4, e4566.
Pushko P, Tretyakova I. 2020. Influenza virus like particles (VLPs): Opportunities for H7N9 vaccine development. Viruses, 12, 518.
Ran X H, Yang Z Y, Bai M Y, Zhang Y, Wen X B, Guo H C, Sun S Q. 2019. Development and validation of a competitive ELISA based on bacterium-original virus-like particles of serotype O foot-and-mouth disease virus for detecting serum antibodies. Applied Microbiology and Biotechnology, 103, 3015–3024.
Sakamoto S, Yusakul G, Tsuneura Y, Putalun W, Usui K, Miyamoto T, Tanaka H, Morimoto S. 2017. Sodium periodate-mediated conjugation of harringtonine enabling the production of a highly specific monoclonal antibody, and the development of a sensitive quantitative analysis method. Analyst, 142, 1140–1148.
Shi J, Deng G H, Kong H H, Gu C Y, Ma S J, Yin X, Zeng X Y, Cui P F, Chen Y, Yang H L, Wan X P, Wang X R, Liu L L, Chen P C, Jiang Y P, Liu J X, Guan Y T, Suzuki Y, Li M, Qu Z Y, et al. 2017. H7N9 virulent mutants detected in chickens in China pose an increased threat to humans. Cell Research, 27, 1409–1421.
Shi J Z, Deng G H, Ma S J, Zeng X Y, Yin X, Li M, Zhang B, Cui P F, Chen Y, Yang H L, Wan X P, Liu L L, Chen P C, Jiang Y P, Guan Y T, Liu J X, Gu W L, Han S Y, Song Y M, Liang L B, et al. 2018. Rapid evolution of H7N9 highly pathogenic viruses that emerged in China in 2017. Cell Host & Microbe, 24, 558–568.
Shien J, Fu L, Wu J, Cheng M, H K, Chang P. 2008. Development of blocking ELISA for detection of antibodies against avian influenza virus of the H7 subtype. Journal of Microbiology, Immunology, and Infection, 41, 369–376.
Skehel J J, Hay A J. 1978. Influenza virus transcription. Journal of General Virology, 39, 1–8.
Stadlbauer D, Amanat F, Strohmeier S, Nachbagauer R, Krammer F. 2018. Cross-reactive mouse monoclonal antibodies raised against the hemagglutinin of A/Shanghai/1/2013 (H7N9) protect against novel H7 virus isolates in the mouse model. Emerging Microbes & Infections, 7, 110.
Sutton T C. 2018. The pandemic threat of emerging H5 and H7 avian influenza viruses. Viruses, 10, 461.
Tong S X, Zhu X Y, Li Y, Shi M, Zhang J, Bourgeois M, Yang H, Chen X F, Recuenco S, Gomez J, Chen L M, Johnson A, Tao Y, Dreyfus C, Yu W L, McBride R, Carney P J, Gilbert A T, Chang J, Guo Z, et al. 2013. New world bats harbor diverse influenza A viruses. PLoS Pathogens, 9, e1003657.
Tu Y C, Chen K Y, Chen C K, Cheng M C, Lee S H, Cheng I C. 2019. Novel application of Influenza A virus-inoculated chorioallantoic membrane to characterize a NP-specific monoclonal antibody for immunohistochemistry assaying. Journal of Veterinary Science, 20, 51–57.
Wang X L, Jiang H, Wu P, Uyeki T M, Feng L, Lai S J, Wang L L, Huo X, Xu K, Chen E F, Wang X X, He J F, Kang M, Zhang R L, Zhang J, Wu J B, Hu S X, Zhang H J, Liu X Q, Fu W J, et al. 2017. Epidemiology of avian influenza A H7N9 virus in human beings across five epidemics in mainland China, 2013–17: An epidemiological study of laboratory-confirmed case series. The Lancet Infectious Diseases, 17, 822–832.
Wang X R, Gu L L, Shi J Z, Xu H F, Zhang Y, Zeng X Y, Deng G H, Li C J, Chen H L. 2017. Development of a real-time RT-PCR method for the detection of newly emerged highly pathogenic H7N9 influenza viruses. Journal of Integrative Agriculture, 16, 2055–2061.
Ward M J, Lycett S J, Avila D, Bollback J P, Leigh Brown A J. 2013. Evolutionary interactions between haemagglutinin and neuraminidase in avian influenza. BMC Evolutionary Biology, 13, 222.
Yang L, Zhu W F, Li X Y, Chen M M, Wu J, Yu P B, Qi S X, Huang Y W, Shi W X, Dong J, Zhao X, Huang W J, Li Z, Zeng X X, Bo H, Chen T, Chen W B, Liu J, Zhang Y, Liang Z L, et al. 2017. Genesis and spread of newly emerged highly pathogenic H7N9 avian viruses in mainland China. Journal of Virology, 91, e01277–e01294.
Yang P, Ma C N, Shi W X, Cui S J, Lu G L, Peng X M, Zhang D T, Liu Y M, Liang H J, Zhang Y, Zhang L, Seale H, Wang Q Y. 2012. A serological survey of antibodies to H5, H7 and H9 avian influenza viruses amongst the duck-related workers in Beijing, China. PLoS ONE, 7, e50770.
Zeng X Y, Chen X H, Ma S J, Wu J J, Bao H M, Pan S X, Liu Y J, Deng G H, Shi J Z, Chen P C, Jiang Y P, Li Y B, Hu J L, Lu T, Mao S G, Guo X F, Liu J L, Tian G B, Chen H L. 2020. Protective efficacy of an H5/H7 trivalent inactivated vaccine produced from Re-11, Re-12, and H7-Re2 strains against challenge with different H5 and H7 viruses in chickens. Journal of Integrative Agriculture, 19, 2294–2300.
Zeng X Y, Tian G B, Shi J Z, Deng G H, Li C J, Chen H L. 2018. Vaccination of poultry successfully eliminated human infection with H7N9 virus in China. Science China (Life Sciences), 61, 1465–1473.
Zhang Q Y, Shi J Z, Deng G H, Guo J, Zeng X Y, He X J, Kong H H, Gu C Y, Li X Y, Liu J X, Wang G J, Chen Y, Liu L L, Liang L B, Li Y Y, Fan J, Wang J L, Li W H, Guan L Z, Li Q M, et al. 2013. H7N9 influenza viruses are transmissible in ferrets by respiratory droplet. Science, 341, 410–414.
Zhang Y, Liu L, Xu H F, Zeng X Y, Shi J Z, Wang X R, Chen H L. 2018. Comparative analysis on HA protein of H7N9 subtype influenza virus with different pathogenicity. China Poultry, 40, 14–17. (in Chinese)
Zhu W F, Yang L, Shu Y L. 2019. Did the highly pathogenic avian influenza A (H7N9) viruses emerged in China raise increased threat to public health? Vector-Borne and Zoonotic Diseases, 19, 22–25.
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