双重实时荧光定量 PCR技术检测中药制剂中 沙门氏菌和大肠埃希菌

中国药物评价 ›› 2022, Vol. 39 ›› Issue (4): 309-313.

双重实时荧光定量 PCR技术检测中药制剂中沙门氏菌和大肠埃希菌

鄢雷娜，陈希，储梅君，刘卫德*

江西省药品检验检测研究院，国家药品监督管理局中成药质量评价重点实验室，江西省药品与医疗器械质量工程技术研究中心，江西南昌 330029

收稿日期:2022-03-23 修回日期:2022-04-07 出版日期:2022-08-28 发布日期:2022-10-08
基金资助:
江西省药品监督管理局科研项目：基于PMA-pPCR技术快速检测口服中药制剂中两种控制菌活菌的研究（2019JS30）

Detection of Salmonella and Escherichia coliin Traditional Chinese Medicine Preparations by Dual Real-time Fluorescence Quantitative PCR

Jiangxi Institute for Drug Control， NMPA Key Laboratory of Quality Evaluation of Traditional Chinese Patent Medicine， Jiangxi Province Engineering Research Center of Drug and Medical Device Quality， Jiangxi Nanchang 330029， China

Received:2022-03-23 Revised:2022-04-07 Online:2022-08-28 Published:2022-10-08

摘要/Abstract

摘要： 目的：建立双重实时荧光定量PCR（multiplex quantitative real-time PCR， multiplex qPCR）快速检测中药制剂中沙门氏菌和大肠埃希菌2种常见控制菌的方法。方法：筛选两种目标菌株的特异性引物与探针，扩增沙门氏菌的invA基因和大肠埃希菌的uidA基因，优化反应体系，建立双重荧光定量PCR方法。结果：两对引物探针对沙门氏菌和大肠埃希菌均有较好地扩增效率，人工污染中药制剂中沙门氏菌的检出限为 101 CFU·mL-1，大肠埃希菌的检出限为101 CFU·mL-1。结论：本研究建立的双重实时荧光定量PCR方法具有特异性强、重复性好，灵敏度高等特点、适用于中药制剂中沙门氏菌和大肠埃希菌的快速检测。

关键词: font-size:medium, ">双重实时荧光定量PCR；中药制剂；快速检测；沙门氏菌；大肠埃希菌

Abstract: Objective：To establish a method for rapid detection of Salmonella and Escherichia coli in traditional Chinese medicine preparations by multiplex quantitative real-time PCR(multiplex qPCR).Methods：The specific primers and probes of two target strains were screened， the invA gene of Salmonella and uidA gene of Escherichia coli were amplified， the reaction system was optimized， and the dual fluorescence quantitative PCR method was established. Results：Two pairs of primer probes can amplify Salmonella and Escherichia coli effectively. The detection limit of Salmonella and Escherichia coli in Artificially Polluted traditional Chinese medicine preparations is 101 CFU·mL-1.Conclusion：The dual real-time fluorescence quantitative PCR method established in this study has the characteristics of strong specificity， good repeatability and high sensitivity. It is suitable for the rapid detection of Salmonella and Escherichia coli in traditional Chinese medicine preparations.

Key words: Dual real-time fluorescence quantitative PCR;Traditional Chinese medicine preparation, Rapid detection;Salmonella;Escherichia coli

中图分类号:

鄢雷娜, 陈希, 储梅君, 刘卫德. 双重实时荧光定量 PCR技术检测中药制剂中沙门氏菌和大肠埃希菌[J]. 中国药物评价, 2022, 39(4): 309-313.

YAN Lei-na, CHEN Xi, CHU Mei-jun, LIU Wei-de. Detection of Salmonella and Escherichia coliin Traditional Chinese Medicine Preparations by Dual Real-time Fluorescence Quantitative PCR[J]. CHINESE JOURNAL OF DRUG EVALUATION, 2022, 39(4): 309-313.

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