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在实验动物健康监测中使用Luminex xTAG检测法同时检测4种原型大鼠细小病毒。

Simultaneous detection of 4 prototypic rat parvoviruses using the luminex xTAG assay in laboratory animal health monitoring.

作者信息

Xu Fengjiao, Yuan Wen, Zhang Tongyuan, Zhu Yujun, Lian Yuexiao, Zhang Yu, Huang Ren, Guo Pengju

机构信息

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.

出版信息

J Virol Methods. 2017 Oct;248:61-65. doi: 10.1016/j.jviromet.2017.05.017. Epub 2017 May 31.

Abstract

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.

摘要

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。

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