National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China; Key Laboratory of Preventive Veterinary Medicine of Hubei Province, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China; Frontiers Science Center for Animal Breeding and Sustainable Production, Wuhan 430070, China.
National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China; Key Laboratory of Preventive Veterinary Medicine of Hubei Province, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China; Frontiers Science Center for Animal Breeding and Sustainable Production, Wuhan 430070, China.
J Virol Methods. 2024 Jul;328:114960. doi: 10.1016/j.jviromet.2024.114960. Epub 2024 May 31.
Canine Infectious Respiratory Disease Complex (CIRDC) is a highly infectious diseases. Canine respiratory coronavirus (CRCoV), Canine influenza virus (CIV), Canine distemper virus (CDV), and Canine parainfluenza virus (CPiV) are crucial pathogens causing CIRDC. Due to the similar clinical symptoms induced by these viruses, differential diagnosis based solely on symptoms can be challenging. In this study, a multiplex real-time PCR assay was developed for detecting the four RNA viruses of CIRDC. Specific primers and probes were designed to target M gene of CRCoV, M gene of CIV, N gene of CDV and NP gene of CPiV. The detection limit is 10 copies/μL for CIV or CRCoV, while the detection limit of CDV or CPiV is 100 copies/μL. Intra-group and inter-group repeatability coefficient of variation (CV) were both less than 2 %. A total of 341 clinical canine samples were analyzed, and the results indicated that the method developed in our study owns a good consistency and better specificity compared with the conventional reverse transcription PCR. This study provides a new method to enable the simultaneous detection of all four pathogens in a single reaction, improving the efficiency for monitoring the prevalence of four viruses in CIRDC, which benefits the control of CIRDC.
犬传染性呼吸道疾病复合症（CIRDC）是一种高度传染性疾病。犬冠状病毒（CRCoV）、犬流感病毒（CIV）、犬瘟热病毒（CDV）和犬副流感病毒（CPiV）是引起 CIRDC 的关键病原体。由于这些病毒引起的临床症状相似，仅基于症状进行鉴别诊断可能具有挑战性。在本研究中，开发了一种用于检测 CIRDC 四种 RNA 病毒的多重实时 PCR 检测方法。针对 CRCoV 的 M 基因、CIV 的 M 基因、CDV 的 N 基因和 CPiV 的 NP 基因设计了特异性引物和探针。CIV 或 CRCoV 的检测限为 10 拷贝/μL，而 CDV 或 CPiV 的检测限为 100 拷贝/μL。组内和组间重复性变异系数（CV）均小于 2%。对 341 份临床犬样本进行分析，结果表明，与常规逆转录 PCR 相比，本研究中建立的方法具有更好的一致性和特异性。本研究提供了一种新方法，可在单个反应中同时检测这四种病原体，提高了监测 CIRDC 中四种病毒流行情况的效率，有助于控制 CIRDC。
