Wan Chunhe, Chen Cuiteng, Cheng Longfei, Chen Hongmei, Fu Qiuling, Shi Shaohua, Fu Guanghua, Liu Rongchang, Huang Yu
Fujian Provincial Key Laboratory for Avian Diseases Control and Prevention, Fujian Animal Diseases Control Technology Development Center, Institute of Animal Husbandry and Veterinary Medicine of Fujian Academy of Agricultural Sciences, Xi-feng Road No.100, Jiantian village, Jin'an district, Fuzhou, 350013, China.
BMC Vet Res. 2018 Sep 3;14(1):267. doi: 10.1186/s12917-018-1600-3.
Muscovy duck parvovirus (MDPV) causes high mortality and morbidity in Muscovy ducks, with the pathogenesis of the virus still unknown in many respects. Specific MDPV detection is often rife with false positive results because of high identity at the genomic nucleotide level and antigenic similarity with goose parvovirus (GPV). The objective of this study was to develop a sensitive, highly specific, and repeatable TaqMan-based real-time PCR (qPCR) assay for facilitating the molecular detection of MDPV.
The specific primers and probe were designed based on the conserved regions within MDPVs, but there was a variation in GPVs of the nonstructural (NS) genes after genetic comparison. After the optimization of qPCR conditions, the detection limit of this qPCR assay was 29.7 copies/μl. The assay was highly specific for the detection of MDPV, and no cross-reactivity was observed with other non-targeted duck-derived pathogens. Intra- and inter-assay variability was less than 2.21%, means a high degree of repeatability. The diagnostic applicability of the qPCR assay was proven that MDPV-positive can be found in cloacal swabs samples, Muscovy duck embryos and newly hatched Muscovy ducklings.
Our data provided incidents that MDPV could be possible vertically transmitted from breeder Muscovy ducks to Muscovy ducklings. The developed qPCR assay in the study could be a reliable and specific tool for epidemiological surveillance and pathogenesis studies of MDPV.
番鸭细小病毒(MDPV)在番鸭中可导致高死亡率和高发病率,该病毒的发病机制在很多方面仍不清楚。由于MDPV与鹅细小病毒(GPV)在基因组核苷酸水平具有高度一致性以及抗原相似性,MDPV的特异性检测常出现大量假阳性结果。本研究的目的是开发一种灵敏、高度特异且可重复的基于TaqMan的实时荧光定量PCR(qPCR)检测方法,以促进MDPV的分子检测。
基于MDPV的保守区域设计了特异性引物和探针,但经过基因比较发现非结构(NS)基因的GPV存在变异。优化qPCR条件后,该qPCR检测方法的检测限为29.7拷贝/微升。该检测方法对MDPV的检测具有高度特异性,未观察到与其他非目标鸭源病原体的交叉反应。批内和批间变异小于2.21%,意味着具有高度可重复性。qPCR检测方法的诊断适用性得到证实,在泄殖腔拭子样本、番鸭胚胎和新孵出的番鸭雏鸭中均能检测到MDPV阳性。
我们的数据提供了MDPV可能从种番鸭垂直传播给番鸭雏鸭的事例。本研究中开发的qPCR检测方法可为MDPV的流行病学监测和发病机制研究提供可靠且特异的工具。