Simultaneous identification of 6 pathogens causing porcine reproductive failure by using multiplex ligation-dependent probe amplification
Simultaneous identification of 6 pathogens causing porcine reproductive failure by using multiplex ligation-dependent probe amplification
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利用多重连接依赖性探针扩增同时鉴定导致猪繁殖失败的 6 种病原体
DOI：
10.1111/tbed.13585
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发表时间：
2020
影响因子：
4.3
通讯作者：
Song Houhui
中科院分区：
文献类型：
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作者：
Zhou Yingshan;Chen Lin;Zhang Lifei;Shao Chunyan;Sun Jing;Jiang Sheng;Song Quanjiang;Zhou Bin;Yang Yongchun;Dong Wanyu;Yang Yang;Wei Fangfang;Fang Weihuan;Wang Xiaodu;Song Houhui
建立了一种多重连接依赖探针扩增（MLPA）方法，用于同时检测猪繁殖与呼吸综合征病毒（PRRSV）、日本脑炎病毒（JEV）、猪瘟病毒（CSFV）、猪圆环病毒2型（PCV 2）、伪狂犬病病毒（PRV）和猪细小病毒（PPV）等6种引起猪繁殖障碍的病毒病原。该检测试剂盒的检测限在6种靶微生物中各不相同，每次MLPA检测试剂盒的检测限为1至8个拷贝。使用346份肝素化猪脐带血标本评价了MLPA检测，并将检测结果与真实的实时PCR结果进行了比较。MLPA检测对PRRSV的特异性和敏感性分别为99.2%和100%;对CSFV、PCV 2、PRV和PPV的特异性和敏感性分别为100%和100%。MLPA检测或真实的时间PCR均未发现样本为JEV阳性。总之，MLPA检测试剂盒的临床灵敏度与真实的实时PCR检测试剂盒相当，为快速筛查猪繁殖障碍相关病毒提供了有用的工具。
We developed a multiplex ligation‐dependent probe amplification (MLPA) assay for the simultaneous detection of 6 clinically relevant viral pathogens causing porcine reproductive failure, that is porcine reproductive and respiratory syndrome virus (PRRSV), Japanese encephalitis virus (JEV), classical swine fever virus (CSFV), porcine circovirus type 2 (PCV2), pseudorabies virus (PRV) and porcine parvovirus (PPV). The limits of detection for the assay varied among the 6 target organisms from 1 to 8 copies per MLPA assay. The MLPA assay was evaluated with 346 heparinized porcine umbilical cord blood specimens, and the results of the assay were compared to those of real‐time PCR. The MLPA assay showed specificities and sensitivities of 99.2% and 100%, respectively, for PRRSV; 100% and 100%, respectively, for CSFV, PCV2, PRV and PPV. No sample was found to be positive for JEV by either the MLPA assay or the real‐time PCR. In conclusion, the MLPA assay has comparable clinical sensitivity to that of real‐time PCR assay and provides a useful tool for fast screening porcine reproductive failure‐associated viruses.