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沙门氏菌LAMP快速检测方法的建立
郭澍强
0
(宁夏检验检疫局检验检疫综合技术中心)
摘要:
为实现沙门氏菌的快速检测,根据沙门氏菌特异性invE、hilA、invA、hut基因编码区序列,分别设计合成4套引物,通过对引物筛选和反应条件优化,建立了以invA特异性基因设计合成为引物的沙门氏菌环介导等温扩增检测方法。本方法对沙门氏菌定性检测灵敏度可达到101CFU/mL(细菌浓度),其灵敏度比普通PCR高10倍且反应结果易于观察。以6种沙门氏菌和8种非沙门氏菌细菌DNA为模板进行LAMP扩增检测,并无交叉反应。结果显示,设计的LAMP引物组和建立的沙门氏菌的LAMP检测方法具有特异性好、灵敏度高、快速、费用低廉等优点,可应用于动物和食品样品中肠道沙门氏菌6个亚种的快速检测。
关键词:  沙门氏菌;LAMP;invA基因  引物;检测方法
DOI:
投稿时间:2019-03-18修订日期:2019-08-02
基金项目:
Development of Rapid LAMP Test Method for Samonella
(Technology Center of Ningxia Entry-exit Inspection and Quarantine Bureau)
Abstract:
In order to establish a rapid detection of Salmonella, four pairs of primers were designed and synthesized according to the invE, hilA, invA and hut gene sequences of Salmonella. By screening primers and optimizing reaction conditions, one Salmonella loop-mediated isothermal amplification assay was developed using invA primers. The sensitivity of the method for qualitative detection of Salmonella could reach 101 CFU/mL, and its sensitivity was 10 times higher than that of ordinary PCR, and the reaction result is easy to observe. LAMP amplification assays were performed when 6 Salmonella and 8 non-Salmonella bacterial DNAs was used as templates. The results showed that the LAMP detection method of Salmonella has the advantages of high specificity, high sensitivity, rapidity and low cost, which can be applied to the rapid detection of six subspecies of Salmonella in animal and food samples.
Key words:  Samonella  LAMP  invA gene  primer  detection method

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