Detection of enterotoxigenic Escherichia coli by real-time polymerase chain reaction by using MGB-TaqMan probe technique / 中华预防医学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To develop a real-time polymerase chain reaction(PCR) based on TaqMan technology by using a new MGB probe for detecting enterotoxigenic Escherichia coli (ETEC) in paper.</p><p><b>METHODS</b>Primers and MGB probe were designed in the ecoding region of heat-stable toxin of ETEC. Real-time PCR detected ETEC by using the exterior standard method with protracting standard curves. The specificity, sensitivity, accuracy, stability of real-time PCR system was evaluated. An internal negative antithesis was added to the real-time PCR system in order to get rid of the false positive of system. Using UNG enzyme expelled the contamination of PCR reaction.</p><p><b>RESULTS</b>Primers and MGB probe were suited to the Real-time PCR. The assay showed that the method was quick, special, sensitive and stable. The real-time PCR system could detect ETEC in a large scale. The assay might be finished in two hour.</p><p><b>CONCLUSION</b>These observations suggested that real-time PCR based on MGB probe should be an excellent candidate for a standard ETEC detection method.</p>