Establishment and application of multiplex PCR for non-O157 H7 STEC virulence genes detection.
- Author:
Xiao-Guang WANG
;
Ying-Hua ZHANG
;
Ping WANG
;
Xiu-Hua CHEN
;
Ling-Fei LUO
;
Yun LIU
;
Ji-Qian LIU
;
Chi-Ping SONG
;
Yang Lin OU
;
Guo-Qiang CHEN
- Publication Type:Journal Article
- MeSH: Escherichia coli Infections; diagnosis; microbiology; Escherichia coli Proteins; genetics; Humans; Multiplex Polymerase Chain Reaction; methods; Shiga-Toxigenic Escherichia coli; genetics; isolation & purification; Virulence Factors; genetics
- From: Chinese Journal of Experimental and Clinical Virology 2013;27(5):388-391
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETraditional 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.
METHODSSix 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.
RESULTSThe 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.
CONCLUSIONThe method we established can be used for non-O157 STEC virulence genes detection and screening with high efficiency and accuracy.