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Indirect ELISA for Detecting Specific IgA antibody to Porcine Epidemic Diarrhea Virus

Lidan Sun, Ligong Chen, Shuangmei Zhou, Yanqin Li, Haiyong Guo, Tanqing Li, Qinye Song


The object of this study is to establish an indirect ELISA method for detecting IgA antibody against porcine epidemic diarrhea virus (PEDV) and provide a technological means to PEDV infection detection and immune effect evaluation. Antigen coating concentration, blocking and dilution buffer, optimal dilutions of test serum and enzyme-labeled antibody were determined and the indirect ELISA method for detecting PEDV-IgA antibody was developed. Furthermore, the specificity, intra- and inter-batch repeatability tests of the ELISA were examined, the coincidence rate were measured between the ELISA and the existing ELISA kit, and the correlation between the specific IgA and the neutralizing antibodies in serum was analyzed. The results showed that the optimal antigen coating concentration of the ELISA was 1μg/mL, the blocking and dilution buffers were PBS containing 5% calf serum and 0.05% Tween-20, and the working concentrations of tested serum and enzyme-labeled antibody were 1:40 and 1:1500, respectively. The method could be used to detect the PEDV specific antibodies without cross reactions with those antibodies against porcine reproductive and respiratory syndrome virus, classical swine fever virus, pseudorabies virus, and porcine circovirus type 2 using the ELISA.The coefficients of variation (CV) of intra- and inter-batch repetitive tests of the ELISA were less than 10%. The coincidence rate was 94.8% when the same serum samples were detected using the ELISA and existing ELISA kit for detecting PEDV antibody. The level of specific IgA antibody was positively correlated with that of the neutralizing antibody in serum (r=0.69, p<0.001).


Porcine Epidemic Diarrhea Virus; S1 Protein; Indirect ELISA; IgA; Neutralizing Antibody; Correlation

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Lee C.Porcine epidemic diarrhea virus: An emerging and re-emerging epizootic swinevirus.Virol J, 2015 12:193.

Li W, Li H, Liu Y, et al. New variants of porcine epidemic diarrhea virus, China, 2011. Emer Infec Dis, 2012, 18(8), 1350-1353.

Sun D, Wang X, Wei S, et al. Epidemiology and vaccine of porcine epidemic diarrhea virus in China: amini-review. J Vet Med Sci, 2016,78(3):355-363.

Pan Y, Tian X, Li W, et al. Isolation and characterization of a variant porcine epidemic diarrhea virus in China. Virol J, 2012, 9:195.

Kocherhans R, Bridgen A, Ackermann M, et al. Completion of the porcine epidemic diarrhea coronavirus (PEDV) genome sequence. Virus Genes, 2001, 23:137-144.

Saif LJ, Pensaert MB, Sestack K, et al. Coronaviruses. In: Zimmerman JJ, Karriker LA, Ramirez A, et al., editors. Diseases of Swine. Ames: Wiley-Blackwell; 2012. p. 501-524.

Jackwood MW, Hilt DA, Callison SA, et al. Spike glycoprotein cleavage recognition site analysis of infectious bronchitis virus. Avian Dis, 2001, 45:366-372.

Wu XD, Shang B, Yang RF, et al. The spike protein of severe acute respiratory syndrome (SARS) is cleaved in virusinfected Vero-E6 cells. Cell Res, 2004, 14(5):400-406.

Li C, Li W, Lucio de Esesarte E, et al. Cell Attachment Domains of the Porcine Epidemic Diarrhea Virus Spike Protein Are Key Targets of Neutralizing Antibodies. J Virol, 2017,91(12). pii: e00273-17.

Sun D, Feng L, Shi H, et al. Identification of two novel B cell epitopes on porcine epidemic diarrhea virus spike protein. Vet Microbiol, 2008, 131(1-2):73-81.

Yamamoto R, Soma J, Nakanishi M, et al.Isolation and experimental inoculation of an S INDEL strain of porcine epidemicdiarrhea virus in Japan. Res Vet Sci, 2015, 103:103-106.

Suzuki T, Shibahara T, Yamaguchi R, et al. Pig epidemic diarrhoea virus S gene variant with a large deletion non-lethal to colostrum-deprived newborn piglets. J Gen Virol, 2016, 97(8):1823-1828.

Hain KS, Joshi LR, Okda F, et al.Immunogenicity of a recombinant parapoxvirus expressing the spike protein of Porcine epidemic diarrhea virus. J Gen Virol, 2016, 97(10):2719-2731.

Gimenez-Lirola LG, Zhang J, Carrillo-Avila JA, et al.Reactivity of Porcine Epidemic Diarrhea Virus Structural Proteins to Antibodiesagainst Porcine Enteric Coronaviruses: Diagnostic Implications[J]. J Clin Microbiol, 2017, 55(5):1426-1436.

Arriba ML, Carvajal A, Pozo J, et al.Mucosal and systemic isotype-specific antibody responses and protection in conventional pigs exposed to virulent or attenuated porcine epidemic diarrheavirus. Vet Immunol Immunopathol,2002, 85(1-2):85-97.

Langel SN, Paim FC, Lager KM, et al.Lactogenic immunity and vaccines for porcine epidemic diarrhea virus (PEDV):Historical and current concepts. Virus Res, 2016, 226:93-107.

Song Q, Stone S, Drebes D, et al. Characterization of anti-porcine epidemic diarrhea virus neutralizing activity inmammary secretions. Virus Res, 2016, 226:85-92.

Gerber PF, Opriessnig T.Detection of immunoglobulin (Ig) A antibodies against porcine epidemic diarrheavirus (PEDV) in fecal and serum samples.MethodsX. 2015, 2:368-373.

BaekPS, Choi HW, Lee S, et al.Efficacy of an inactivated genotype 2b porcine epidemic diarrhea virus vaccine inneonatal piglets. Vet Immunol Immunopathol, 2016, 174:45-49.

Goede D, Murtaugh MP, Nerem J, et al..Previous infection of sows with a "mild" strain of porcine epidemic diarrheavirus confers protection against infection with a "severe" strain. Vet Microbiol, 2015, 176(1-2):161-164.

Ding Zhenjiang, Ku Xugang, Yan Guiwei, et al. Establishment of indirect ELISA for detection of IgA antibody in swine epidemic diarrhea. Animal Husbandry and Veterinary Medicine, 2014. 46(11):1-5.

Yao Zuojun, Hao Daren, Bai Yun, etc. Detection of porcine epidemic diarrhea virus S1 Indirect ELISA Method. Journal of Animal Husbandry and Veterinary Medicine, 2017, 48, 6, 1085-1091.

Hua Yao, Grace Wai Wong, Li Yu, etc. " Prokaryotic Expression and Indirect Expression of Major Antigen Epitope Region of S and ELISA of Porcine Epidemic Diarrhea Virus, Establishment of Detection Method, Microbiological bulletin, 2016,43 (2): 434-443.

Gerber PF, Gong Q, Huang YW, et al. Detection of antibodies against porcine epidemic diarrhea virus in serum andcolostrum by indirect ELISA. Vet J, 2014, 202(1):33-36.

KnuchelM, Ackermann M, Müller HK, et al. An ELISA for detection of antibodies against porcine epidemic diarrhoea virus (PEDV) based on the specific solubility of the viral surface glycoprotein. Vet Microbiol, 1992, 32(2):117-134.

Wang X, Wang Z, Xu H, et al. Orally Administrated Whole Yeast Vaccine Against Porcine Epidemic Diarrhea Virus Induced High Levels of IgA Response in Mice and Piglets. Viral Immunol, 2016, 29(9):526-531.

Zhang Y, Zhang X, Liao X, et al. Construction of a bivalent DNA vaccine co-expressing S genes of transmissible gastroenteritis virus and porcine epidemic diarrhea virus delivered by attenuated Salmonella typhimurium Virus Genes, 2016, 52(3):354-364.

Cao L, Ge X, Gao Y, et al. Putative phage-display epitopes of the porcine epidemic diarrhea virus S1 protein and their anti-viral activity. Virus Genes, 2015, 51(2):217-224.

PoonsukK, Giménez-Lirola LG, Zhang J, et al.Does Circulating Antibody Play a Role in the Protection of Piglets againstPorcine Epidemic Diarrhea Virus? PLoS One, 2016, 11(4):e0153041.



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