一种检测乙型脑炎病毒抗体阻断ELISA的建立及其初步应用

doi:10.11843/j.issn.0366-6964.2018.09.017

Abstract

Abstract:

Japanese encephalitis is a mosquito transmitted zoonosis caused by Japanese encephalitis virus (JEV) that seriously harms the health of human and animals, such as pigs and horses. In order to establish a specific method for detection the JEV antibody in various animal serum, a blocking ELISA method was established using recombinant EDⅢ protein and horseradish peroxidase (HRP) conjugated JEV specific Monoclonal antibody 1B10. Results were as follows:The percentage of interruption rate (PI) of 100 JEV antibody negative serum samples were statistically analyzed, the cut-off of blocking ELISA was determined as the sample presenting a calculated PI ≥ 34% were considered positive; while samples with a calculated PI ≤ 25% were rated negative; and those presenting PI between 25% and 34% were considered inconclusive. No cross-reaction with antibody positive serum against PCV2, PRV, CSFV, FMDV and PRRSV was observed in the test. And the intra-and inter-assay variation were less than 5%. Compared with the commercialization JEV antibody detection ELISA kit, the agreement rates of the new established blocking ELISA was 97.2%. The specificity and sensitivity of the blocking ELISA were 98.5% and 94.3%, respectively. The blocking ELISA was used to test against 515 serum samples collected from swine, cattle and sheep, among which 74.5%, 13.3% and 9.2% of samples were JEV specific antibody-positive. The detection results of randomly selected swine, cattle and sheep by the serum neutralization test were agree with blocking ELISA. As a result, a blocking ELISA was successfully established for detecting JEV antibody of swine, cattle and sheep in the present study.

Key words: Japanese encephalitis virus, envelope protein, HRP-labeled MAb, blocking ELISA

CLC Number:

S852.659.6

JIANG Ya, NIU Ji-chen, NI Hui, WEI Dong-mei, ZHOU Guo-dong, CAO Rui-bing. Development and Preliminary Application of a Blocking ELISA for Detection of Japanese Encephalitis Virus Antibody[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(9): 1961-1968.

References

[1] 陈溥言. 兽医传染病学[M]. 5版. 北京:中国农业出版社, 2006.
CHEN P Y. Veterinary lemology[M]. 5th ed. Beijing:China Agriculture Press, 2006. (in Chinese)
[2] MORITA K. Molecular epidemiology of Japanese encephalitis in East Asia[J]. Vaccine, 2009, 27(50):7131-7132.
[3] WANG H Y, TAKASAKI T, FU S H, et al. Molecular epidemiological analysis of Japanese encephalitis virus in China[J]. J Gen Virol, 2007, 88(3):885-894.
[4] CHAMBERS T J, HAHN C S, GALLER R, et al. Flavivirus genome organization, expression, and replication[J]. Annu Rev Microbiol, 1990, 44:649-688.
[5] YE Q, XU Y P, ZHANG Y, et al. Genotype-specific neutralization determinants in envelope protein:implications for the improvement of Japanese encephalitis vaccine[J]. J Gen Virol, 2015, 96(8):2165-2175.
[6] LUCA V C, ABIMANSOUR J, NELSON C A, et al. Crystal structure of the Japanese encephalitis virus envelope protein[J]. J Virol, 2012, 86(4):2337-2346.
[7] TURTLE L, SOLOMON T. Japanese encephalitis-the prospects for new treatments[J]. Nat Rev Neurol, 2018, 14(5):298-313.
[8] CSANK T, DRZEWNIOKOVÁ P, KORYTÁR L, et al. A serosurvey of flavivirus infection in horses and birds in slovakia[J]. Vector Borne Zoonotic Dis, 2018, 18(4):206-213.
[9] PIYASENA T B H, SETOH Y X, HOBSON-PETERS J, et al. Differential diagnosis of flavivirus infections in horses using viral envelope protein domain Ⅲ antigens in enzyme-linked immunosorbent assay[J]. Vector Borne Zoonotic Dis, 2017, 17(12):825-835.
[10] GALULA J U, CHANG G J J, CHUANG S T, et al. Establishment of an algorithm using prM/E-and NS1-specific IgM antibody-capture enzyme-linked immunosorbent assays in diagnosis of Japanese encephalitis virus and west nile virus infections in humans[J]. J Clin Microbiol, 2016, 54(2):412-422.
[11] 祖立闯,王金良,管宇,等. 流行性乙型脑炎病毒E蛋白的截短表达与间接ELISA诊断方法的建立[J]. 中国预防兽医学报, 2010, 32(3):200-204, 231.
ZU L C, WANG J L, GUAN Y, et al. Development of an indirect ELISA for detecting antibodies to japanese encephalitis virus using recombinant truncated glycoprotein E expressed in E. coil[J]. Chinese Journal of Preventive Veterinary Medicine, 2010, 32(3):200-204, 231. (in Chinese)
[12] HIROTA J, SHIMOJI Y, SHIMIZU S. New sensitive competitive enzyme-linked immunosorbent assay using a monoclonal antibody against nonstructural protein 1 of west nile virus NY99[J]. Clin Vaccine Immunol, 2012, 19(2):277-283.
[13] 倪慧,王钰,毛岩,等. 日本脑炎病毒囊膜蛋白单克隆抗体的制备及特性分析[J]. 畜牧与兽医, 2016, 48(7):1-9.
NI H, WANG Y, MAO Y, et al. Preparation and characterization of monoclonal antibodies against envelope protein of Japanese encephalitis virus[J]. Animal Husbandry & Veterinary Medicine, 2016, 48(7):1-9. (in Chinese)
[14] FRIGUET B, DJAVADI-OHANIANCE L, PAGES J, et al. A convenient enzyme-linked immunosorbent assay for testing whether monoclonal antibodies recognize the same antigenic site. Application to hybridomas specific for the β₂-subunit of Escherichia coli tryptophan synthase[J]. J Immunol Methods, 1983, 60(3):351-358.
[15] HOMBACH J, SOLOMON T, KURANE I, et al. Report on a WHO consultation on immunological endpoints for evaluation of new Japanese encephalitis vaccines, WHO, Geneva, 2-3 September, 2004[J]. Vaccine, 2005, 23(45):5205-5211.
[16] KEDL R M, KAPPLER J M, MARRACK P. Epitope dominance, competition and T cell affinity maturation[J]. Curr Opin Immunol, 2003, 15(1):120-127.
[17] 马淑娟,熊益权,张琼花,等. 三种ELISA试剂盒检测猪流行性乙型脑炎病毒血清IgG抗体比较[J]. 中国人兽共患病学报, 2015, 31(5):423-426.
MA S J, XIONG Y Q, ZHANG Q H, et al. Comparison of three commercial ELISA kits for detection of antibodies against Japanese encephalitis virus in swine sera[J]. Chinese Journal of Zoonoses, 2015, 31(5):423-426. (in Chinese)
[18] MAO L, LI W L, ZHOU T C, et al. Development of a blocking ELISA for Caprine parainfluenza virus type 3[J]. J Virol Methods, 2017, 250:59-65.
[19] CHAI C X, WANG Q, CAO S J, et al. Serological and molecular epidemiology of Japanese encephalitis virus infections in swine herds in China, 2006-2012[J]. J Vet Sci, 2018, 19(1):151-155.
[20] RICKLIN M E, GARCÍA-NICOLÍS O, BRECHBUHL D, et al. Vector-free transmission and persistence of Japanese encephalitis virus in pigs[J]. Nat Commun, 2016, 7:10832.
[21] MANSFIELD K L, HERNÁNDEZ-TRIANA L M, BANYARD A C, et al. Japanese encephalitis virus infection, diagnosis and control in domestic animals[J]. Vet Microbiol, 2017, 201:85-92.
[22] HUBÁLEK Z, RUDOLF I, NOWOTNY N. Arboviruses pathogenic for domestic and wild animals[J]. Adv Virus Res, 2014, 89:201-275.
[23] CHOWDHURY S, KHAN S U, CRAMERI G, et al. Serological evidence of henipavirus exposure in cattle, goats and pigs in Bangladesh[J]. PLoS Negl Trop Dis, 2014, 8(11):e3302.

Development and Preliminary Application of a Blocking ELISA for Detection of Japanese Encephalitis Virus Antibody

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