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Establishment of the HDA method of detecting Escherichia coli O157:H7

Yang Da-wei

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Abstract

In this paper,a new method,which based on helicase-dependent isothermal DNA amplification(HDA) to detect Escherichia coli O157:H7 of sample,was established.A highly specific set of primers was syn-thesized to target the rfbE gene of Escherichia coli O157:H7 so as to establish helicase-dependent isothermal DNA amplification method.Specific and sensitivity were tested compared with PCR method.The results showed that the sensitivity of HDA was 4.3×103 cfu·mL-1 which was similar to the result of PCR method.Detecting Es-cherichia coli O157:H7 with HDA is specific and sensitive as well as with PCR method and has lower instrumen-tal requirement.

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What this paper is about

In this paper,a new method,which based on helicase-dependent isothermal DNA amplification(HDA) to detect Escherichia coli O157:H7 of sample,was established.A highly specific set of primers was syn-thesized to target the rfbE gene of Escherichia coli O157:H7 so as to establish helicase-dependent isothermal DNA amplification method.Specific and sensitivity were tested compared with PCR method.The results showed that the sensitivity of HDA was 4.3×103 cfu·mL-1 which was similar to the result of PCR method.Detecting Es-cherichia coli O157:H7 with HDA is specific and sensitive as well as with PCR method and has lower instrumen-tal requirement.

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Available abstract

In this paper,a new method,which based on helicase-dependent isothermal DNA amplification(HDA) to detect Escherichia coli O157:H7 of sample,was established.A highly specific set of primers was syn-thesized to target the rfbE gene of Escherichia coli O157:H7 so as to establish helicase-dependent isothermal DNA amplification method.Specific and sensitivity were tested compared with PCR method.The results showed that the sensitivity of HDA was 4.3×103 cfu·mL-1 which was similar to the result of PCR method.Detecting Es-cherichia coli O157:H7 with HDA is specific and sensitive as well as with PCR method and has lower instrumen-tal requirement.

Key concepts: Escherichia coli, Molecular biology, DNA, Biology, Loop-mediated isothermal amplification, Gene, Polymerase chain reaction, Genetics

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