Establishment of a multiplex RT-PCR assay for identification of atmospheric virus contamination in pig farms
Establishment of a multiplex RT-PCR assay for identification of atmospheric virus contamination in pig farms
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养猪场大气病毒污染多重RT-PCR检测方法的建立
DOI：
10.1016/j.envpol.2019.07.028
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发表时间：
2019-10-01
影响因子：
8.9
通讯作者：
Liu, Xingyou
中科院分区：
文献类型：
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作者：
Li, Han;Wei, Xiaobing;Liu, Xingyou
病原体在猪场的传播不仅会将疾病传染给其他猪甚至在猪场工作的农民，还会对猪场周围居民的生活大气环境造成污染。因此，有必要建立一种快速、简便的监测方法。本研究通过对猪场常见病毒全基因组序列的分析，结合引物设计、反应参数优化，建立了一种能同时鉴定猪瘟病毒（CSFV）、乙型脑炎病毒（JEV）、猪繁殖与呼吸综合征病毒（PRRSV）、猪圆环病毒2型（PCV-2）、猪伪狂犬病病毒（PRV）和猪细小病毒（PPV），它们在养猪场中很常见。该方法的最低可检测浓度为10（-3）ng/μ L，具有高度特异性。应用多重RT-PCR方法对我国不同城市4个猪场的空气样品进行检测。结果与单PCR结果一致。因此，本研究有望为重大病毒的预警机制和疾病防控体系提供必要的技术支持。(C)2019爱思唯尔有限公司版权所有。
Spread of pathogens in pig farms not only causes transfection of diseases to other pigs or even farmers working in the farms, but also induces pollution to the living atmospheric environment of the residents around the farm. Therefore, it is necessary to establish a rapid and simple monitoring method. In this study, full genome sequences of common viruses were analyzed in pig farms, in combination with the design of primers, optimization of the reaction parameters, so as to establish a multiplex RT-PCR assay for the identification of classical swine fever virus (CSFV), Japanese encephalitis virus (JEV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus Type 2 (PCV-2), porcine pseudorabies virus (PRV) and porcine parvovirus virus (PPV), which are common in pig farms. This method has a minimal detectable concentration of 10(-3) ng/mu L, which is highly specific. Furthermore, multiplex RT-PCR was applied to examine air samples from 4 pig farms located in different cities of China. The results were in line with those obtained by single PCR. Therefore, this study can be expected to provide essential technique support for the early warning mechanism as well as disease prevention and control system against the major viruses. (C) 2019 Elsevier Ltd. All rights reserved.