Detection of IPaH1 gene in the Shigella and optimization of PCR conditions

Objective To detect the expression of IPaH1 genes in Shigella and PCR conditions optimized.Methods The expression of ipaH gene levels in Shigella boydii,wild Shigella,Escherichia coli,Enterohemorrhage E.coli(O157:H7),Salmonella enterica subsp,Enterobacter sakazakii,Proteus vulgaris,Bacillus cereus was determined by PCR,and the optimization of PCR conditions on annealing temperature and primer concentration was evoluated.Results ① IPaH1 gene specific expression were detected in Shigella while no expression were detected in Escherichia coli,Enterohemorrhage E.coli(O157:H7),Salmonella enterica subsp,Enterobacter sakazakii,Proteus vulgaris,Bacillus cereus,Enterococcus faecalis,Staphylococcus aureus,Listeria monocytogenes,Vibrio parahaemolyticus,Clostridium perfringens;②Amplification of Shigella ipaH1 gene by PCR,the first primers of IPaH1-1 optimal annealing temperature is 55.0℃-59.0℃,and the second ones of IPaH1-2 is 51.0℃-55.0℃;the final optimal concentration of primers is 0.1 μM.Conclusion IPaH1 gene specifically expressed in Shigella and its optimal annealing temperature was respectively 56°C,52°C,the optimal final concentration of primers is 0.1 μM.