印第安纳沙门菌环介导等温扩增检测方法的建立

doi:10.11843/j.issn.0366-6964.2021.02.029

畜牧兽医学报 ›› 2021, Vol. 52 ›› Issue (2): 560-564.doi: 10.11843/j.issn.0366-6964.2021.02.029

• 研究简报 • 上一篇

印第安纳沙门菌环介导等温扩增检测方法的建立

龚建森¹, 徐敬潇¹, 付立霞², 张笛¹, 董永毅³, 徐步¹, 窦新红^1*

1. 中国农业科学院家禽研究所, 扬州 225125;
2. 扬州大学动物科学与技术学院, 扬州 225009;
3. 江苏省动物疫病预防控制中心, 南京 210036

收稿日期:2020-07-20 出版日期:2021-02-23 发布日期:2021-02-24
通讯作者: 窦新红,主要从事家禽疫病防控及健康养殖模式研究,E-mail:douxinhong611@163.com
作者简介:龚建森(1980-),男,江苏扬州人,博士,主要从事禽病防治研究,E-mail:jjsensen@163.com,Tel:0514-85599282
基金资助:
国家自然科学基金（31772758）；江苏现代农业产业技术体系建设项目（JATS〔2018〕250；JATS〔2019〕345）

Establishment of a Loop-Mediated Isothermal Amplification Method for Rapid Detection of Salmonella Indiana

GONG Jiansen¹, XU Jingxiao¹, FU Lixia², ZHANG Di¹, DONG Yongyi³, XU Bu¹, DOU Xinhong^1*

1. Poultry Institute, Chinese Academy of Agricultural Sciences, Yangzhou 225125, China;
2. College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China;
3. Jiangsu Animal Disease Prevention and Control Center, Nanjing 210036, China

Received:2020-07-20 Online:2021-02-23 Published:2021-02-24

摘要/Abstract

摘要： 旨在解决印第安纳沙门菌传统检测方法的缺陷，建立快速、特异、灵敏的分子检测方法。通过比较基因组学方法获取印第安纳沙门菌特异性检测靶标，进一步设计用于环介导等温扩增的引物组。通过对反应条件优化，建立针对印第安纳沙门菌的特异性分子检测方法。结果显示，该检测方法特异性强、耗时短，检测下限为59拷贝·反应^-1。应用该方法对临床分离菌株进行检测，结果不仅与国家标准（GB4789.4—2016）检测结果一致，还可用于自凝菌株检测。本研究建立的方法能实现印第安纳沙门菌的快速检测，将在临床诊断中发挥重要作用。

关键词: 印第安纳沙门菌, 环介导等温扩增技术, 比较基因组学, 检测

Abstract: The objective of the present study was to establish a molecular method for rapid detection of Salmonella Indiana and overcome the shortcomings of traditional detection method rely on selective isolation, biochemical identification and serological typing. In this study, comparative genomics was used to identify the serospecific gene for S. Indiana, and a set of primers for LAMP was further designed. The specific molecular detection method for S. Indiana was established by optimizing the reaction conditions. The results showed that the method was specific and time-consuming, and the detection limit was 59 copies per reaction. The results of this method are consistent with GB4789.4—2016, and can also be used for the detection of self-coagulating bacteria. The detection method established in this study can rapid detection of S. Indiana, and will play an important role in the clinical diagnosis of this pathogen.

Key words: Salmonella Indiana, loop-mediated isothermal amplification, comparative genomics, detection

中图分类号:

S852.61

龚建森, 徐敬潇, 付立霞, 张笛, 董永毅, 徐步, 窦新红. 印第安纳沙门菌环介导等温扩增检测方法的建立[J]. 畜牧兽医学报, 2021, 52(2): 560-564.

GONG Jiansen, XU Jingxiao, FU Lixia, ZHANG Di, DONG Yongyi, XU Bu, DOU Xinhong. Establishment of a Loop-Mediated Isothermal Amplification Method for Rapid Detection of Salmonella Indiana[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(2): 560-564.

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印第安纳沙门菌环介导等温扩增检测方法的建立

Establishment of a Loop-Mediated Isothermal Amplification Method for Rapid Detection of Salmonella Indiana

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