[Study of rapid method on enterotoxigenic Escherichia coli detected by real-time fluoresence quantitative PCR].
OBJECTIVE
To develop a real-time PCR for detecting enterotoxigenic Escherichia coli (ETEC) based on TaqMan technology.


METHODS
Primers and probes were designed in the coding region of heat-stable enterotoxin, heat-labile enterotoxin of ETEC. ETEC were detected by real-time fluoresence quantitative PCR, making use of the exterior standard curve which was described by several different concentration. The speciality, sensitivity, accuracy, repetition, and stability of real-time fluoresence quantitative PCR system were evaluated.


RESULTS
Primers and TaqMan probes were suited to the real-time fluoresence quantitative PCR. The assay showed that the method could be rapid, special, sensitive and stabile. The real-time PCR system could detect ETEC between 10(0)-10(7) DNA copies/reaction. The assay should be finished in two hours.


CONCLUSIONS
It was suggested that real-time fluoresence quantitative PCR based on TaqMan probe could be a rapid, sensitive and special method. It is significant that the excellent method could control diarrhoea caused by ETEC.
Dai
Juan
J
Lab Medicine Department, Chengdu Medical College, Chengdu 610083, China. daijuan_2004_2007@126.com
Li
Yufeng
Y
Yang
Xiao
X
Yuan
Lixing
L
D054307:Enterotoxigenic Escherichia coli / Q000235:genetics / Q000302:isolation & purification*; D005456:Fluorescent Dyes; D015345:Oligonucleotide Probes / Q000737:chemistry / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D005456:Fluorescent Dyes; D015345:Oligonucleotide Probes