鸭圆环病毒、鸭坦布苏病毒、麝香鸭呼肠孤病毒和新型鸭呼肠孤病毒多重 qPCR 检测方法的建立与应用。

Development and application of a multiplex qPCR assay for the detection of duck circovirus, duck Tembusu virus, Muscovy duck reovirus, and new duck reovirus.

机构信息

Guangxi Center for Animal Disease Control and Prevention, Nanning, 530001, China.

College of Animal Science and Technology, Guangxi University, Nanning, 530005, China.

出版信息

Virus Genes. 2023 Feb;59(1):91-99. doi: 10.1007/s11262-022-01946-0. Epub 2022 Oct 18.

DOI:10.1007/s11262-022-01946-0

PMID:36258144

Abstract

A multiplex qPCR assay was developed to simultaneously detect duck circovirus (DuCV), duck Tembusu virus (DTMUV), Muscovy duck reovirus (MDRV), and novel duck reovirus (NDRV), but it did not amplify other viruses, including duck virus enteritis (DVE), infectious bursal disease virus (IBDV), avian reovirus (ARV), H5 avian influenza virus (H5 AIV), H7 avian influenza virus (H7 AIV), H9 avian influenza virus (H9 AIV), Newcastle disease virus (NDV), and Muscovy duck parvovirus (MDPV), and the detection limit for DuCV, DTMUV, MDRV, and NDRV was 1.51 × 10 copies/μL. The intra- and interassay coefficients of variation were less than 1.54% in the repeatability test with standard plasmid concentrations of 1.51 × 10, 1.51 × 10, and 1.51 × 10 copies/μL. The developed multiple qPCR assay was used to examine 404 clinical samples to verify its practicability. The positivity rates for DuCV, DTMUV, MDRV, and NDRV were 26.0%, 9.9%, 4.0%, and 4.7%, respectively, and the mixed infection rates for DuCV + DTMUV, DuCV + MDRV, DuCV + NDRV, MDRV + NDRV, DTMUV + MDRV, and DTMUV + NDRV were 2.7%, 1.2%, 1.2%, 1.0%, 0.5%, and 0.7%, respectively.

摘要

建立了一种多重 qPCR 检测方法，用于同时检测鸭圆环病毒（DuCV）、鸭坦布苏病毒（DTMUV）、Muscovy 鸭呼肠孤病毒（MDRV）和新型鸭呼肠孤病毒（NDRV），但不能扩增其他病毒，包括鸭病毒性肠炎（DVE）、传染性法氏囊病病毒（IBDV）、禽呼肠孤病毒（ARV）、H5 禽流感病毒（H5 AIV）、H7 禽流感病毒（H7 AIV）、H9 禽流感病毒（H9 AIV）、新城疫病毒（NDV）和 Muscovy 鸭细小病毒（MDPV），DuCV、DTMUV、MDRV 和 NDRV 的检测限为 1.51×10 拷贝/μL。在标准质粒浓度为 1.51×10、1.51×10 和 1.51×10 拷贝/μL 的重复性试验中，内和间试验的变异系数均小于 1.54%。该多重 qPCR 检测方法用于检测 404 份临床样本，以验证其实用性。DuCV、DTMUV、MDRV 和 NDRV 的阳性率分别为 26.0%、9.9%、4.0%和 4.7%，DuCV+DTMUV、DuCV+MDRV、DuCV+NDRV、MDRV+NDRV、DTMUV+MDRV 和 DTMUV+NDRV 的混合感染率分别为 2.7%、1.2%、1.2%、1.0%、0.5%和 0.7%。