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投稿时间：2020-07-16
投稿时间：2020-07-16
中文摘要: 该研究建立多重酶链式反应法快速检测鼠伤寒沙门氏菌3 种毒力基因的检测方法。以鼠伤寒沙门氏菌毒力基因mgtC、inva、mogA、sseL、araB 设计合成特异性引物，提取鼠伤寒沙门氏菌的DNA 为扩增模板，通过单重聚合酶链式反应技术（polymerase chain reaction，PCR）验证引物特异性，利用引物特异性、Tm 值等综合因素通过多重聚合酶链式反应技术（multiple polymerase chain reaction，MPCR）检测方法实现沙门氏菌多种毒力基因的检测，并优化MPCR 试验参数，提高检测灵敏性。结果表明，在多对特异性引物中进行MPCR 反应，筛选出mgtC、mogA、araB 3 种毒力基因进行MPCR 反应的特异性强、无交叉污染，且该方法能检测的最低灵敏度为0.0165 ng/μL。成功建立快速检测食品中沙门氏菌3 种毒力基因的方法，该方法检测特异性强、灵敏度高，检测限低。
中文关键词: 沙门氏菌 毒力基因 多重聚合酶链式反应技术 快速检测 特异性强
Abstract:To establish a multiple enzyme chain reaction method for rapid detection of three virulence genes of Salmonella typhimurium.Specific primers were designed and synthesized using the virulence genes mgtC，inva，mogA， sseL and araB of Salmonella typhimurium. The DNA of Salmonella typhimurium was extracted as the amplification template.The primer specificity was verified by single-plex polymerase chain reaction（PCR）.The primer specificity，Tm value and other comprehensive factors to realize the detection of multiple virulence genes of Salmonella through MPCR detection methods， and optimize the MPCR experimental parameters to improve the detection sensitivity. Results showed that MPCR reactions were carried out on multiple pairs of specific primers， and the three virulence genes of mgtC， mogA， and araB were screened for specificity and no crosscontamination. The minimum sensitivity of this method was 0.0165 ng/μL. The method for rapid detection of three virulence genes of Salmonella in food has been successfully established. The method has strong specificity，high sensitivity and low detection limit.
keywords: Salmonella virulence genes multiplex polymerase chain reaction rapid detection strong specificity
文章编号：202112027 中图分类号： 文献标志码：
基金项目:天津市科技计划项目（18YFZCNC01140）
| Author Name | Affiliation |
| TAN Yan，XIAO Zi-ming，LI Xiao-lin，ZHANG Jie，SONG Wen-jun，WEI Ji-ping，QIAO Chang-sheng | School of Biotechnology and Food Science，Tianjin University of Commerce，Tianjin 300134，China |
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