title: [Establishment and application of multiplex PCR for non-O157 H7 STEC virulence genes detection].
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 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.
  
  
  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.
  
  
  CONCLUSIONS
  The method we established can be used for non-O157 STEC virulence genes detection and screening with high efficiency and accuracy.
authors.forename: Xiao-Guang
authors.initials: XG
authors.lastname: Wang
authors.forename: Ying-Hua
authors.initials: YH
authors.lastname: Zhang
authors.forename: Ping
authors.initials: P
authors.lastname: Wang
authors.forename: Xiu-Hua
authors.initials: XH
authors.lastname: Chen
authors.forename: Ling-Fei
authors.initials: LF
authors.lastname: Luo
authors.forename: Yun
authors.initials: Y
authors.lastname: Liu
authors.forename: Ji-Qian
authors.initials: JQ
authors.lastname: Liu
authors.forename: Chi-Ping
authors.initials: CP
authors.lastname: Song
authors.forename: Yang Lin
authors.initials: YL
authors.lastname: Ou
authors.forename: Guo-Qiang
authors.initials: GQ
authors.lastname: Chen
mesh_terms: D004927:Escherichia coli Infections / Q000175:diagnosis / Q000382:microbiology*; D029968:Escherichia coli Proteins / Q000235:genetics; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D054323:Shiga-Toxigenic Escherichia coli / Q000235:genetics / Q000302:isolation & purification*; D037521:Virulence Factors / Q000235:genetics*
chemical_list: D029968:Escherichia coli Proteins; D037521:Virulence Factors