Rapid Detection and Identification of Listeria Monocytogenes Proficiency Testing Samples by TaqMan Real-Time Fluorescence PCR
Rapid screening detection and identification of Vibrio parahaemolyticus proficiency validation samples based on TaqMan fluorescence PCR.In order to verify the accuracy of the method,according to the GB 4789.7-2013 National Food Safety Standard Food Microbiology Test-Vibrio parahaemolyticus Test for traditional culture detection,through enrichment,isolation,purification to obtain the pure colonies of suspected target bacteria,through preliminary identification and identification for research and judgment,to determine the identification of the use of domestic dried biochemical identification kit,API20E reagent strips and VITEK2 automatic microbial identification system.At the same time,as the traditional culture method,the genomic DNA in the aqueous culture of 3％sodium chloride peptone and the colony genomic DNA on the isolation plate were extracted,specific Taqman primers and fluorescent probes were designed,and the rapid screening and identification technology of real-time fluorescence PCR technology was established.The results showed that the six biochemical tests in the dried biochemical kit were consistent with the standards,but there were few identification test items,and the results needed to be subjectively determined.After the identification of 21 biochemical tests in the API20E kit,sample 071 was found to be Vibrio parahaemolyticus by the bacterial library software,with an identification percentage of 98.6％and a T value of 1.Detected and analyzed by VITEK2 automatic microbial identification system,Vibrio parahaemolyticus was detected in sample 071,and Salmonella enteritidis and Escherichia coli were detected in sample 276.Samples 071 were detected by fluorescence PCR identification of suspicious bacteria on 3％sodium chloride peptone aqueous culture and TCBS,and sample 276 samples of Vibrio haemolyticus were not detected.Real-time fluorescence PCR is consistent with the standard test results in the rapid screening of bacterial enrichment solution and the rapid identification of suspicious colonies,and can be used as an auxiliary detection and identification method for various complex samples.
万方数据
维普
