Rapid detection of enteric pathogens multiple PCR System.doc

 PAGE \* MERGEFORMAT 10 Rapid detection of enteric pathogens multiple PCR System Salmonella, Shigella, Escherichia coli, Enterotoxigenic Escherichia coli is a common cause of human and animal food poisoning, dysentery and bacterial intestinal pathogens, serious harm to human health and livestock safety 〔 1〕 . Therefore, the establishment of a rapid, simple, accurate and specific detection method of great significance. The current testing methods for common pathogens detected the existence of a long cycle, heavy workload, the required reagents are numerous and false-negative rate, sensitivity is low, or false-positive rate, poor specificity shortcomings. PCR technology provides an ideal detection method. In this study, multiplex PCR method can be detected in the same reaction system in different bacteria, the whole process requires only 2 ~ 3h, application of a bright future. 1 Materials and methods 1  1 experimental strain of Salmonella enteritidis, Salmonella Abo Ding, Ji Chu Salmonella Derby Salmonella typhimurium sand bacteria, Shigella flexneri 2a, Shigella flexneri 2b, Song Shigella bacteria, acid citrate edge Bacillus, Proteus, river bacteria, Staphylococcus aureus, Bacillus subtilis (Sichuan Provincial Center for Disease Control and Prevention); enterotoxigenic Escherichia coli C83921, enterotoxigenic Escherichia coli C83902 (China monitored by veterinary medicine); invasion Escherichia coli (China Biological Products Institute). 1  2 reagent dye, buffer solution, TaqDNA polymerase, dNTPs, nucleic acid dyes (GV), agarose, Marker, etc. (Beijing Parkson Cypriot Genentech). 1  3 major equipment PCR gene amplification device, desktop high-speed centrifuges, UV-visible gel imaging system, nucleic acid analyzers, micro-plus kind of gun (Germany Eppendorf Company); electrophoresis tank, DYY-2 regulator Steady Flow Electrophoresis (Beijing 61 Instrument Factory). 1  4 Method 1  4  1 primer design Salmonella histidine transport operon gene frag