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韩国导致猪繁殖失败的病毒病原体的流行情况,以及多重实时 PCR 和 RT-PCR 检测方法的开发。

Prevalence of viral agents causing swine reproductive failure in Korea and the development of multiplex real-time PCR and RT-PCR assays.

机构信息

Animal Disease Diagnostic Division, Animal and Plant Quarantine Agency, Gimcheon, 39660, Republic of Korea.

Mediandiagnostics, Chuncheon, 24399, Gangwon-do, Republic of Korea.

出版信息

Biologicals. 2024 May;86:101763. doi: 10.1016/j.biologicals.2024.101763. Epub 2024 Apr 18.

Abstract

This study aimed to investigate the prevalence of viral agents causing reproductive failure in pigs in Korea. In addition, two types of multiplex real-time PCR (mqPCR) were developed for the simultaneous detection of Aujeszky's disease virus (ADV) and porcine parvovirus (PPV) in mqPCR and encephalomyocarditis virus (EMCV) and Japanese encephalitis virus (JEV) in reverse transcription mqPCR (mRT-qPCR). A total of 150 aborted fetus samples collected from 2020 to 2022 were analyzed. Porcine reproductive and respiratory syndrome virus was the most prevalent (49/150 32.7%), followed by porcine circovirus type 2 (31/150, 20.7%), and PPV1 (7/150, 4.7%), whereas ADV, EMCV, and JEV were not detected. The newly developed mqPCR and mRT-qPCR could simultaneously detect and differentiate with high sensitivities and specificities. When applied to aborted fetuses, the newly developed mqPCR for PPV was 33.3% more sensitivities than the previously established diagnostic method. Amino acid analysis of the VP2 sequences of PPV isolates revealed considerable similarity to the highly pathogenic Kresse strain. This study successfully evaluated the prevalence of viral agents causing reproductive failure among swine in Korea, the developed mqPCR and mRT-qPCR methods could be utilized as effective and accurate diagnostic methods for the epidemiological surveillance of ADV, PPV, EMCV, and JEV.

摘要

本研究旨在调查韩国导致猪繁殖失败的病毒病原体的流行情况。此外,还开发了两种多重实时 PCR(mqPCR)方法,用于 mqPCR 中同时检测猪繁殖与呼吸综合征病毒(PRRSV)和猪细小病毒(PPV),以及 mRT-qPCR 中检测脑心肌炎病毒(EMCV)和日本脑炎病毒(JEV)。共分析了 2020 年至 2022 年期间收集的 150 份流产胎儿样本。猪繁殖与呼吸综合征病毒最为普遍(49/150,32.7%),其次是猪圆环病毒 2 型(31/150,20.7%)和 PPV1(7/150,4.7%),而 ADV、EMCV 和 JEV 未检出。新开发的 mqPCR 和 mRT-qPCR 可同时以高灵敏度和特异性进行检测和区分。当应用于流产胎儿时,新开发的用于检测 PPV 的 mqPCR 比先前建立的诊断方法的灵敏度高 33.3%。对 PPV 分离株的 VP2 序列进行氨基酸分析显示,其与高致病性 Kresse 株具有相当大的相似性。本研究成功评估了韩国导致猪繁殖失败的病毒病原体的流行情况,开发的 mqPCR 和 mRT-qPCR 方法可作为 ADV、PPV、EMCV 和 JEV 的流行病学监测的有效和准确的诊断方法。

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