Establishment and application of multiplex PCR for non-O157 H7 STEC virulence genes detection / 中华实验和临床病毒学杂志

ABSTRACT

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>Six virulence genes of non-O157: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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>The method we established can be used for non-O157 STEC virulence genes detection and screening with high efficiency and accuracy.</p>