[1]肖盛中,马振国,李 岩,等.基于重组Erns蛋白的牛病毒性腹泻病毒间接ELISA检测方法的建立[J].中国预防兽医学报,2019,(03):267-272.[doi:0.3969/j.issn.1008-0589.201806039]
 XIAO Sheng-zhong,MA Zhen-guo,LI Yan,et al.Development of an indirect ELISA for detection of antibody against bovine viral diarrhea virus by using the recombinant Erns[J].Chinese journal of preventive veterinary medicine,2019,(03):267-272.[doi:0.3969/j.issn.1008-0589.201806039]
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基于重组Erns蛋白的牛病毒性腹泻病毒间接ELISA检测方法的建立()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2019年03
页码:
267-272
栏目:
诊断和检测技术
出版日期:
2019-04-25

文章信息/Info

Title:
Development of an indirect ELISA for detection of antibody against bovine viral diarrhea virus by using the recombinant Erns
文章编号:
1008-0589(2019)03-0267-06
作者:
 

肖盛中1马振国1李 岩1马旭升1李 爽2杨若松2盛金良1*

 (1. 石河子大学 动物科技学院,新疆 石河子 832003;2. 北京维德维康生物技术有限责任公司,北京 100095)
Author(s):
 

XIAO Sheng-zhong1 MA Zhen-guo1 LI Yan1 MA Xu-sheng1 LI Shuang2YANG Ruo-song2 SHENG Jin-liang1*

 (1. College of Animal Science and Technology, Shihezi University, Shihezi 832003, China;
2. Weideweikang Biotech Company, Beijing 100095, China)
关键词:
牛病毒性腹泻粘膜病毒Erns蛋白间接ELISA
Keywords:
bovine viral diarrhea virus  Erns  indirect ELISA
分类号:
S852.65
DOI:
0.3969/j.issn.1008-0589.201806039
文献标志码:
A
摘要:
为建立中牛病毒性腹泻病毒(BVDV)抗体间接ELISA方法,本研究对新疆地区多个发病牛场临床发病牛肛拭子提取总RNA,RT-PCR扩增BVDV-Erns基因截短片段(681 bp),将该片段克隆于表达载体中,经大肠杆菌表达并纯化后作为ELISA包被抗原,建立了间接ELISA检测方法。特异性试验显示该方法与牛其它常见5种病原阳性血清均无交叉反应。敏感性试验显示在阳性血清1∶1 600倍稀释时仍能够检出。重复性试验显示批内、批间变异系数均小于10 %。与爱德士(IDEXX)商品化试剂盒相比较,总符合率较高。应用该方法对新疆地区2个散户牛场和3个规模化牛场的564份牛血清样品进行了检测,结果显示BVDV抗体阳性率分别为11.76 %、13.04 %、82.76 %、77.96 %、75.82 %,表明BVDV感染在新疆部分地区已呈高发态势。该方法的建立对新疆地区BVDV的血清流行病学调查和养牛业的健康发展具有重要意义。
 
Abstract:
In order to establish an assay for detection of the antibodies against bovine viral diarrhea virus (BVDV), the total RNA was extracted from the anal swab of clinically ill cattle in Xinjiang, and the 681bp truncated fragment of BVDV Erns gene was amplified by RT-PCR, and the indirect ELISA was developed with the recombinant Erns expressed in E.coli as the coating antigen. The results of specificity test showed that the detection method specifically reacted with the BVDV positive serum, but had no cross-reaction with the positive sera against other 5 kinds of pathogens of cattle. The results of sensitivity test indicated that the detection limit for positive serum was 1∶1,600. The results of reproducibility test showed that the coefficient of intra- and inter-variations were both less than 10%. Compared with the IDEXX kit for detection of BVDV antibodies, the total positive rate of the indirect ELISA developed in this study was very high. Moreover, a total of 564 serum samples collected from 2 retail cattle farms and 3 large-scale cattle farms were tested by the established ELISA and the positive rates of BVDV were 11.76%, 13.04%, 82.76%, 77.96% and 75.82%, respectively. The method was of great practical significance for detection of BVDV antibodies in cattle farms and for the healthy development of the cattle industry.

参考文献/References:

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(本文编辑:李 爽)

更新日期/Last Update: 2019-04-26