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应用逆转录重组酶聚合酶扩增检测方法诊断牛冠状病毒的新方法。

A new approach for diagnosis of bovine coronavirus using a reverse transcription recombinase polymerase amplification assay.

机构信息

Department of Botany and Microbiology, College of Science, King Saud University, 11451 Riyadh, Saudi Arabia; Department of Virology, Faculty of Veterinary Medicine, Cairo University, 12211 Giza, Egypt.

出版信息

J Virol Methods. 2013 Nov;193(2):337-40. doi: 10.1016/j.jviromet.2013.06.027. Epub 2013 Jun 28.

Abstract

Bovine coronavirus (BCoV) is an economically significant cause of calf scours and winter dysentery of adult cattle, and may induce respiratory tract infections in cattle of all ages. Early diagnosis of BCoV helps to diminish its burden on the dairy and beef industry. Real-time RT-PCR assay for the detection of BCoV has been described, but it is relatively expensive, requires well-equipped laboratories and is not suitable for on-site screening. A novel assay, using reverse transcription recombinase polymerase amplification (RT-RPA), for the detection of BCoV is developed. The BCoV RT-RPA was rapid (10-20 min) and has an analytical sensitivity of 19 molecules. No cross-reactivity with other viruses causing bovine gastrointestinal and/or respiratory infections was observed. The assay performance on clinical samples was validated by testing 16 fecal and 14 nasal swab specimens and compared to real-time RT-PCR. Both assays provided comparable results. The RT-RPA assay was significantly more rapid than the real-time RT-PCR assay. The BCoV RT-RPA constitutes a suitable accurate, sensitive and rapid alternative to the common measures used for BCoV diagnosis. In addition, the use of a portable fluorescence reading device extends its application potential to use in the field and point-of-care diagnosis.

摘要

牛冠状病毒(BCoV)是引起犊牛腹泻和成年牛冬季痢疾的重要经济原因,也可能引起各年龄段牛的呼吸道感染。BCoV 的早期诊断有助于减轻其对奶牛和肉牛产业的负担。已描述了用于检测 BCoV 的实时 RT-PCR 检测方法,但它相对昂贵,需要设备齐全的实验室,不适合现场筛选。本研究开发了一种用于检测 BCoV 的新型检测方法,即逆转录重组酶聚合酶扩增(RT-RPA)。BCoV 的 RT-RPA 快速(10-20 分钟),分析灵敏度为 19 个分子。与引起牛胃肠道和/或呼吸道感染的其他病毒无交叉反应。通过检测 16 份粪便和 14 份鼻拭子标本,对临床样本的检测性能进行了验证,并与实时 RT-PCR 进行了比较。两种检测方法均提供了可比的结果。RT-RPA 检测方法比实时 RT-PCR 检测方法明显更快。BCoV RT-RPA 是一种合适的替代方法,与用于 BCoV 诊断的常用方法相比,具有准确性、敏感性和快速性。此外,使用便携式荧光读数设备扩展了其在现场和即时诊断中的应用潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8665/7113639/ded3ab4f7bee/gr1.jpg

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