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  • Title: [Detection of enterotoxigenic Escherichia coli by real-time polymerase chain reaction by using MGB-TaqMan probe technique].
    Author: Dai J, Li YF, Yuan LX, Yang X.
    Journal: Zhonghua Yu Fang Yi Xue Za Zhi; 2008 Feb; 42(2):103-6. PubMed ID: 18642662.
    Abstract:
    OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSION: These observations suggested that real-time PCR based on MGB probe should be an excellent candidate for a standard ETEC detection method.
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