Kim Jong-Min, Kim Hye-Ryung, Jeon Gyu-Tae, Baek Ji-Su, Kwon Oh-Deog, Park Choi-Kyu
Animal Disease Intervention Center, College of Veterinary Medicine, Kyungpook National University, Daegu 41566, Republic of Korea.
Animals (Basel). 2023 Feb 8;13(4):598. doi: 10.3390/ani13040598.
Two species of porcine parainfluenza viruses (PPIV), PPIV1 and PPIV5, are globally distributed in pig herds and associated with porcine respiratory diseases, and a diagnostic tool for the simultaneous detection of the two viruses is required. In this study, a TaqMan probe-based duplex real-time reverse transcription polymerase chain reaction (dqRT-PCR) assay was first developed for the differential detection of PPIV1 and PPIV5 nucleocapsid protein (NP) genes in porcine clinical samples. The dqRT-PCR assay was highly sensitive, its limit of detection was approximately 10 RNA copies/reaction, it specifically amplified the targeted NP genes of PPIV1 and PPIV5 without cross-reacting with other porcine pathogens, and their clinical detection rates were 15.2% and 0.7%, respectively. The results from 441 clinical samples taken from 278 Korean domestic pig farms showed that the prevalence of PPIV1 and PPIV5 was 11.2% and 1.1%, respectively, and co-infection of both viruses was confirmed in a farm, suggesting that PPIV1 and PPIV5 are co-circulating in current Korean pig herds. Phylogenetic analysis based on the partial NP genes suggested that genetically diverse PPIV1 strains are circulating in Korean pig herds. The developed dqRT-PCR assay was found to be an accurate, reliable, and quantitative detection tool for PPIV1 and PPIV5 RNA in clinical pig samples and will be useful for etiological and epidemiological studies and the control of viral infections in the field.
两种猪副流感病毒（PPIV），即PPIV1和PPIV5，在全球猪群中广泛分布并与猪呼吸道疾病相关，因此需要一种同时检测这两种病毒的诊断工具。在本研究中，首次开发了一种基于TaqMan探针的双重实时逆转录聚合酶链反应（dqRT-PCR）检测方法，用于鉴别检测猪临床样本中的PPIV1和PPIV5核衣壳蛋白（NP）基因。该dqRT-PCR检测方法具有高度敏感性，其检测限约为10个RNA拷贝/反应，能特异性扩增PPIV1和PPIV5的靶向NP基因，不与其他猪病原体发生交叉反应，其临床检出率分别为15.2%和0.7%。从278个韩国国内猪场采集的441份临床样本结果显示，PPIV1和PPIV5的流行率分别为11.2%和1.1%，在一个猪场中确认了两种病毒的共同感染，这表明PPIV1和PPIV5在当前韩国猪群中共同传播。基于部分NP基因的系统发育分析表明，遗传多样的PPIV1毒株在韩国猪群中传播。已开发的dqRT-PCR检测方法被发现是一种用于临床猪样本中PPIV1和PPIV5 RNA的准确、可靠且定量的检测工具，将有助于病因学和流行病学研究以及该领域病毒感染的防控。
