流行性出血病多克隆抗体C-ELISA检测方法的建立

doi:10.11843/j.issn.0366-6964.2018.07.013

Abstract

Abstract:

The objective of this research was to establish a simple and fast operation method for detecting epizootic haemorrhagic disease virus(EHDV) antibodies. In this research, a competitive ELISA(C-ELISA) method was developed, by using purified EHDV-6 coated ELISA plates and guinea pig anti-EHDV-2 antibody as a competitive antibody. The assay had a high specificity and reproducibility, with optimal dilution of the multi-clone antibody at 1:10 000 and virus antigen at 5.12 μg·mL^-1(1:10 000). The critical value of the detection method was 50%, through the serum testing of cows and sheep, including negative and positive 270 copies each. Specific tests showed that the ELISA method can only detect different serotypes of EHDV antibodies, with better group specificity. Positive serum tests on known antibody titers showed that the established C-ELISA method was more sensitive than the serum neutralization test(SNT) and Agar gel immunodiffusion test(AGID). While the serum and anticoagulant samples collected from monitoring animals were detected by C-ELISA, SNT, RT-PCR and virus isolation respectively, the results of ELISA were consistent with the other two methods. These results indicate that the C-ELISA is a rapid, sensitive and stable method for the detection of EHDV antibodies.

Key words: epizootic haemorrhagic disease virus (EHDV), multi-clone antibody, competitive ELISA (C-ELISA)

CLC Number:

S854.43

ZHU Jian-bo, YANG Zhen-xing, XIAO Lei, GAO Lin, MIAO Hai-sheng, HE Yu-wen, MENG Jin-xin, YANG Heng, LI Hua-chun. Establishment of a Rapid Competitive ELISA for Detecting Antibodies against Epizootic Haemorrhagic Disease Virus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(7): 1440-1450.

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Establishment of a Rapid Competitive ELISA for Detecting Antibodies against Epizootic Haemorrhagic Disease Virus

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