Development of a quadruple PCR-based gene microarray for detection of vaccine and wild-type classical swine fever virus, African swine fever virus and atypical porcine pestivirus.
Development of a quadruple PCR-based gene microarray for detection of vaccine and wild-type classical swine fever virus, African swine fever virus and atypical porcine pestivirus.
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开发基于四重PCR的基因微阵列，用于检测疫苗和野生型猪瘟病毒、非洲猪瘟病毒和非典型猪瘟病毒
DOI：
10.1186/s12985-022-01933-9
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发表时间：
2022-11-29
期刊：
影响因子：
4.8
通讯作者：
Zhang, Qian-yi
中科院分区：
文献类型：
--
作者：
Xia, Ying-ju;Xu, Lu;Zhao, Jun-jie;Li, Yuan-xi;Wu, Rui-zhi;Song, Xiang-peng;Zhao, Qi-zu;Liu, Ye-bing;Wang, Qin;Zhang, Qian-yi
关键词：
猪瘟、非洲猪瘟和非典型猪瘟是目前困扰中国养猪业的急性、强毒和传染性的病毒性疾病，可导致仔猪木乃伊或死胎，导致生猪死亡。为了更好地预防、控制和消除中国的这些病毒性疾病，除了检测猪瘟病毒和猪细小病毒外，迫切需要猪瘟病毒感染和疫苗接种的诊断分析。本研究建立了一种基于四重聚合酶链式反应的基因芯片技术，根据CSFV和APPV的保守区，同时检测CSFV野毒株和疫苗株。对42份实验室确认的样品，包括其他10种猪病毒病的阳性样品，用该方法进行了检测，以确认其高特异性。本方法对猪瘟野毒和疫苗株的检出限分别为6.98和6.92拷贝/µL，检出限分别为2.56 × 10和1.8 × 10拷贝/微克L。与标准逆转录聚合酶链式反应或定量聚合酶链式反应检测猪瘟病毒(GB/T 26875-2018年)、猪瘟病毒(MARR第172期)或猪细小病毒(CN108611442A型)比较，符合率为100%。结果表明，该方法具有较高的灵敏度，能特异性地区分ASFV、APPV和CSFV，包括CSFV感染和免疫。该方法为这三种病毒的快速检测和准确诊断提供了一种实用、简便、经济、可靠的检测方法，在这三种疾病的流行病学调查、预防和控制与消除中具有良好的应用前景。
Classical swine fever (CSF), African swine fever (ASF), and atypical porcine pestivirus (APPV) are acute, virulent, and contagious viral diseases currently hampering the pig industry in China, which result in mummification or stillbirths in piglets and mortality in pigs. Diagnostic assays for the differentiation of infection and vaccination of CSFV, in addition to the detection of ASFV and APPV, are urgently required for better prevention, control, and elimination of these viral diseases in China. A quadruple PCR-based gene microarray assay was developed in this study to simultaneously detect wild-type and vaccine CSFV strains, ASFV and APPV according to their conserved regions. Forty-two laboratory-confirmed samples, including positive samples of 10 other swine viral diseases, were tested using this assay to confirm its high specificity. This assay's limit of detections (LODs) for the wild-type and vaccine CSFV were 6.98 and 6.92 copies/µL. LODs for ASFV and APPV were 2.56 × 10 and 1.80 × 10 copies/µL, respectively. When compared with standard RT-PCR or qPCR for CSFV (GB/T 26875–2018), ASFV (MARR issue No.172), or APPV (CN108611442A) using 219 clinical samples, the coincidence was 100%. The results showed that this assay with high sensitivity could specifically distinguish ASFV, APPV, and CSFV, including CSFV infection and immunization. This assay provides a practical, simple, economic, and reliable test for the rapid detection and accurate diagnosis of the three viruses and may have good prospects for application in an epidemiological investigation, prevention, and control and elimination of these three diseases.
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