牛棒状杆菌环介导等温扩增快速检测方法的建立及初步应用

doi:10.3969/j.issn.1674-5817.2019.05.003

实验动物与比较医学 ›› 2019, Vol. 39 ›› Issue (5): 349-355.DOI: 10.3969/j.issn.1674-5817.2019.05.003

牛棒状杆菌环介导等温扩增快速检测方法的建立及初步应用

冯洁^1,2, 张泉¹, 钱淼¹, 谢建云², 高诚²

1.扬州大学兽医学院, 扬州 225009;
2.上海实验动物研究中心, 上海市实验动物质量监督检验站, 上海 201203

收稿日期:2019-04-29 出版日期:2019-10-25 发布日期:2021-01-05
作者简介:冯洁(1981-), 女, 副研究员, 主要从事实验动物质量控制研究。E-mail: moyifj@163.com
基金资助:
上海市科技创新行动计划(17140900600)

Development of a Loop-mediated Isothermal Amplification Assay for Detection of Corynebacterium bovis

FENG Jie^1,2, ZHANG Quan¹, QIAN Miao¹, XIE Jian-yun², GAO Cheng²

1. College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China;
2. Shanghai Laboratory Animal Research Center, Shanghai Quality Monitoring Center of Laboratory Animals, Shanghai 201203, China

Received:2019-04-29 Online:2019-10-25 Published:2021-01-05

摘要/Abstract

摘要： 目的建立牛棒状杆菌(C.bovis)环介导等温扩增(LAMP)快速检测方法。方法根据牛棒状杆菌16S rRNA基因保守区域设计一组特异性引物,对反应条件进行优化并对特异性和敏感性进行分析。结果建立的LAMP扩增方法仅能检测出牛棒状杆菌,其他常见细菌未见特异性扩增,检出牛棒状杆菌的最低模板量为118 fg/μL。结论建立的牛棒状杆菌LAMP检测方法操作简单、快速高效,结果易于观察,对仪器设备要求低,可用于牛棒状杆菌的快速检测。

关键词: 牛棒状杆菌(C.bovis), 环介导等温扩增(LAMP), 检测

Abstract: Objective To establish a rapid and specific loop-mediated isothermal amplification (LAMP) assay for Corynebacterium bovis(C.bovis). Methods A set of specific primers were designed based on the 16S rRNA of C. Bovis. The reaction parameters of LAMP assay were optimized and the specificity and sensitivity were determined. Results Among the tested bacteria, only C.bovis could be detected by the LAMP assay, no other bacteria were positive. The detection limit of the LAMP assay was 118 fg/μL genomic DNA of C. bovis. Conclusions The detection could be completed without expensive equipment and it was simple, specific, sensitive, and the result was ready to read-out. LAMP could be a good alternative method for a rapid detection and surveillance of C. bovis.

Key words: Corynebacterium bovis (C.bovis), Loop-mediated isothermal amplification (LAMP), Detection

中图分类号:

Q95-33

冯洁, 张泉, 钱淼, 谢建云, 高诚. 牛棒状杆菌环介导等温扩增快速检测方法的建立及初步应用[J]. 实验动物与比较医学, 2019, 39(5): 349-355.

FENG Jie, ZHANG Quan, QIAN Miao, XIE Jian-yun, GAO Cheng. Development of a Loop-mediated Isothermal Amplification Assay for Detection of Corynebacterium bovis[J]. Laboratory Animal and Comparative Medicine, 2019, 39(5): 349-355.

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牛棒状杆菌环介导等温扩增快速检测方法的建立及初步应用

Development of a Loop-mediated Isothermal Amplification Assay for Detection of Corynebacterium bovis

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