实时荧光定量PCR法检测中药制剂中沙门菌

Detection of Salmonella in Traditional Chinese Medicine Preparations by Real-time Fluorescence Quantitative PCR

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。

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.