Establishment and application of multiplex PCR for non-O157 H7 STEC virulence genes detection

VernacularTitle:非O157型产志贺毒素大肠埃希菌毒力基因多重PCR检测方法的建立与应用
Keywords: Escherichia coli; Genes; Polymerase chain reaction
From: Chinese Journal of Experimental and Clinical Virology 2013;27(5):388-391
CountryChina
Language:Chinese
Abstract: Objective Traditional detection approaches for non-O157 STEC are both time and labour consuming in diseases surveillance.Virulence genes detection based on multiplex PCR could not only improve the detection efficiency but also increase the accuracy.Methods Six virulence genes of nonO157:H7 (stx1,stx2,eae,hly,etpD,katP6) were detected by two groups of trebling PCRs.The multiplex PCRs were optimized by melting curve analysis in SYBR Green Ⅰ real-time PCR.Testing result of multiplex PCR was consistent with serological testing.Results The sensitivity limits of the multiplex PCR for stx1,stx2,eaeP,etpD,katP,and hly were 10 ng/ml,120 ng/ml,110 ng/ml,165 ng/ml,85 ng/ml,and 15 ng/ml,respectively,which is similar with that of single PCR.When the multiplex PCR was applied in 120 adults and 90 children diarrhea samples detection,13 cases were detected for non-O157 positive.Conclusion The method we established can be used for non-O157 STEC virulence genes detection and screening with high efficiency and accuracy.