Detection of Staphylococcus aureus enterotoxin A gene by RT-PCR

Staphylococcus aureus has been considered as one of the most important bacteria causing bacterial food poison.At home and abroad,there are many cases which are caused by Staphylococcus aureus enterotoxin.The conventional method of inspecting food for possible contamination by coagulase positive Staphylococcus aureus enterotoxin is time-consuming,which would take 3-5 d to complete,besides that it is not sensitive enough to detect Staphylococcus aureus enterotoxin.In this study,a rapid method was established to detect Staphyloco-ccus aureus enterotoxin in food to shorten the time of the detection method,increased the sensitivity of method and provide a technical help,which based on the sensitivity and specificity of RT-PCR.The total RNA was isolated from Staphylococcus aureus respectively by hot phenol method and TRIZol method,and these two methods were compared.The mRNA was catalyzed by reverse transcriptase to produce cDNA,and sea was amplified by PCR using specific primers for the Staphylococcus aureus enterotoxin A gene.After that the para-meters of PCR were optimized to procure plenty of positive target genes.The conclusion of this research showed that the effect of hot phenol method is better than TRIZol method in preparation of RNA,which upgrades the cycle counts and time of reaction to develop a method of detection on Staphylococcus aureus enterotoxin A gene by RT-PCR.