Xu Fengjiao, Yuan Wen, Zhang Tongyuan, Zhu Yujun, Lian Yuexiao, Zhang Yu, Huang Ren, Guo Pengju
Guangdong Laboratory Animals Monitoring Institute, Guangzhou 510640, China; Guangdong Provincial Key Laboratory of Laboratory Animals, Guangzhou 510640, China.
Guangdong Laboratory Animals Monitoring Institute, Guangzhou 510640, China.
J Virol Methods. 2017 Oct;248:61-65. doi: 10.1016/j.jviromet.2017.05.017. Epub 2017 May 31.
There are currently four rat parvoviruses including Kilham rat virus (KRV), Toolans H-1 parvovirus (H-1virus), rat parvovirus type 1a (RPV-1a) and rat minute virus (RMV). Virus detection methods are commonly based on conventional PCR - agarose gel electrophoresis or serological assay methods These methods are both time-consuming and lack specificity. In this study, we developed a bead array xTAG assay for the simultaneous detection and discrimination of four rat parvoviruses. The detection limits ranged from 100 to 1000 copies/μL of input purified plasmid DNA. We examined 50 clinical specimens and 15 facal samples by xTAG assay and conventional PCR. The results showed a high consistency except for several weak positive infections. It demonstrated that the xTAG-multiplex PCR method is specific, sensitive and suitable for high throughput platforms for rat parvovirus screening of clinical samples and contaminated biological materials.
目前有四种大鼠细小病毒,包括基尔汉姆大鼠病毒(KRV)、图兰斯H-1细小病毒(H-1病毒)、1a型大鼠细小病毒(RPV-1a)和大鼠微小病毒(RMV)。病毒检测方法通常基于常规PCR-琼脂糖凝胶电泳或血清学检测方法。这些方法既耗时又缺乏特异性。在本研究中,我们开发了一种微珠阵列xTAG检测法,用于同时检测和区分四种大鼠细小病毒。检测限为每微升输入的纯化质粒DNA 100至1000拷贝。我们通过xTAG检测法和常规PCR检测了50份临床标本和15份粪便样本。结果显示,除了几例弱阳性感染外,一致性较高。这表明xTAG多重PCR方法具有特异性、敏感性,适用于临床样本和受污染生物材料的大鼠细小病毒高通量筛选平台。