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基于重组衣壳蛋白的间接酶联免疫吸附测定法用于水貂圆环病毒感染检测的开发与应用

Development and application of an indirect enzyme-linked immunosorbent assay based on recombinant capsid protein for the detection of mink circovirus infection.

作者信息

Ge Junwei, Cui Xingyang, Shi Yunjia, Zhao Lili, Wei Chengwei, Wen Shanshan, Xia Shuang, Chen Hongyan

机构信息

College of Veterinary Medicine, Northeast Agricultural University, Harbin, 150030, China.

Northeastern Science Inspection Station, China Ministry of Agriculture Key Laboratory of Animal Pathogen Biology, Harbin, 150030, China.

出版信息

BMC Vet Res. 2018 Jan 26;14(1):29. doi: 10.1186/s12917-018-1337-z.

Abstract

BACKGROUND

Mink circovirus (MiCV) is a newly discovered pathogen associated with mink diarrhea. The prevalence and economic importance of this virus remain poorly understood, and no specific serological assay has been developed for the diagnosis of MiCV infection.

RESULTS

In this study, a recombinant capsid protein antigen expressed in Escherichia coli was utilized to establish an indirect enzyme-linked immunosorbent assay (iELISA). Results revealed that the assay had no cross-reactivity with other related pathogens, and the respective sensitivity and specificity of the proposed iELISA were 92.31% and 91.67% compared with those obtained of Western blot on 138 serum samples from minks. The correlation coefficient between iELISA and Western blot was 0.838 (p > 0.05). iELISA was applied to detect MiCV antibodies in 683 clinical serum samples from different farms from the major mink industry province in China, and 21 of 24 farms with 163 of 683 (23.87%) individuals were tested positive for MiCV antibodies. The positive rates of each of the 21 flocks ranged from 2.33% to 73.68%.

CONCLUSIONS

These results indicated that iELISA was a sensitive and specific method suitable for the large-scale detection of MiCV infections in mink. This study provided an effective method for the serological diagnosis and positive rate investigation of MiCV infection.

摘要

背景

水貂圆环病毒(MiCV)是一种新发现的与水貂腹泻相关的病原体。人们对该病毒的流行情况和经济重要性仍知之甚少,且尚未开发出用于诊断MiCV感染的特异性血清学检测方法。

结果

在本研究中,利用在大肠杆菌中表达的重组衣壳蛋白抗原建立了间接酶联免疫吸附测定(iELISA)。结果显示,该检测方法与其他相关病原体无交叉反应,与对138份水貂血清样本进行的蛋白质免疫印迹法相比,所提出的iELISA的敏感性和特异性分别为92.31%和91.67%。iELISA与蛋白质免疫印迹法之间的相关系数为0.838(p>0.05)。iELISA应用于检测来自中国主要水貂养殖省份不同养殖场的683份临床血清样本中的MiCV抗体,24个养殖场中有21个养殖场的683只水貂中有163只(23.87%)检测出MiCV抗体呈阳性。21个鸡群中每个鸡群的阳性率在2.33%至73.68%之间。

结论

这些结果表明,iELISA是一种灵敏且特异的方法,适用于大规模检测水貂中的MiCV感染。本研究为MiCV感染的血清学诊断和阳性率调查提供了一种有效的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5e1f/5787310/606bf2917919/12917_2018_1337_Fig1_HTML.jpg

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