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引用本文:刘卫德,张文婷,易巧,熊骏,刘绪平.实时荧光定量PCR法检测中药制剂中沙门菌[J].中国现代应用药学,2021,38(7):836-840.
LIU Weide,ZHANG Wenting,YI Qiao,XIONG Jun,LIU Xuping.Detection of Salmonella in Traditional Chinese Medicine Preparations by Real-time Fluorescence Quantitative PCR[J].Chin J Mod Appl Pharm(中国现代应用药学),2021,38(7):836-840.
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实时荧光定量PCR法检测中药制剂中沙门菌
刘卫德, 张文婷, 易巧, 熊骏, 刘绪平
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, 南昌 330029
摘要:
目的 采用实时荧光定量PCR技术,建立一种中药制剂中沙门菌的快速检测方法。方法 将中药制剂加入适量的胰酪大豆胨液体培养基中进行增菌后,取增菌液以热裂解法提取总DNA,采用实时荧光定量PCR法特异性检测沙门菌。结果 建立了中药制剂中沙门菌的实时荧光定量PCR检测法,检测可在24 h内完成,灵敏度达每10 g(10 mL)供试品1 cfu,特异性为100%,采用该方法检测18批次样品结果与药典方法一致。结论 采用实时荧光定量PCR法可明显缩短中药制剂中沙门菌检测周期,具有良好的灵敏度和特异性,可作为药典检测方法的有效补充。
关键词:  中药制剂  沙门氏菌  实时荧光定量PCR  快速检测
DOI:10.13748/j.cnki.issn1007-7693.2021.07.011
分类号:R917
基金项目:江西省食品药品监督管理局科研项目(2017YX05);江西省重点研发计划项目(20192BBGL70052);江西省药品监督管理局科研项目(2019JS30)
Detection of Salmonella in Traditional Chinese Medicine Preparations by Real-time Fluorescence Quantitative PCR
LIU Weide, ZHANG Wenting, YI Qiao, XIONG Jun, LIU Xuping
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, Nanchang 330029, China
Abstract:
OBJECTIVE To establish a rapid detect method for Salmonella in traditional Chinese medicine by real-time fluorescence quantitative PCR. METHODS Adding the traditional Chinese medicine to an appropriate amount of trypticase soy broth medium for bacteria enrichment, and the total bacterial DNA in the enrichment fluid was extracted by boiling method, then the real-time fluorescence quantitative PCR method was used to specifically detect the Salmonella in the sample. RESULTS A real-time fluorescence quantitative PCR assay for the detection of Salmonella in traditional Chinese medicine was established. The detection could be completed within 24 h, sensitivity was up to per 10 g(10 mL) samples 1 cfu, and the specificity was 100%. 18 batches of samples were detected by this method and the results were identical with detected by the method described in Chinese Pharmacopoeia. CONCLUSION For Real-time fluorescence quantitative PCR method, the detection time of Salmonella in traditional Chinese medicine preparations can significantly shorten and with good sensitivity and specificity, which can be an effective supplement of the test method included in Chinese Pharmacopoeia.
Key words:  traditional Chinese medicine preparations  Salmonella  real-time fluorescence quantitative PCR  rapid analysis
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