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Establishment of the Helicase-dependent isothermal DNA amplification method of detecting Listeria monocytogenes

Yang Da-wei

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Abstract

In this paper,a new method for detection of Listeria monocytogenes was established based on helicase-dependent isothermal DNA amplification(HDA).A pair of primers to target the iap gene(AF532297) of L.monocytogenes was synthesized and the amplification was conducted under 65℃ for 90 minutes after initial denature and annealing to amplify a 105 bp product from L.monocytogenes,but not from other bacteria.The results showed that the sensitivity of HDA was 7.8×103 cfu/mL which was similar to the result of PCR method.Therefore,the rapid and easy-to-use method for detecting L.monocytogenes has a potential application.

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What this paper is about

In this paper,a new method for detection of Listeria monocytogenes was established based on helicase-dependent isothermal DNA amplification(HDA).A pair of primers to target the iap gene(AF532297) of L.monocytogenes was synthesized and the amplification was conducted under 65℃ for 90 minutes after initial denature and annealing to amplify a 105 bp product from L.monocytogenes,but not from other bacteria.The results showed that the sensitivity of HDA was 7.8×103 cfu/mL which was similar to the result of PCR method.Therefore,the rapid and easy-to-use method for detecting L.monocytogenes has a potential application.

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Available abstract

In this paper,a new method for detection of Listeria monocytogenes was established based on helicase-dependent isothermal DNA amplification(HDA).A pair of primers to target the iap gene(AF532297) of L.monocytogenes was synthesized and the amplification was conducted under 65℃ for 90 minutes after initial denature and annealing to amplify a 105 bp product from L.monocytogenes,but not from other bacteria.The results showed that the sensitivity of HDA was 7.8×103 cfu/mL which was similar to the result of PCR method.Therefore,the rapid and easy-to-use method for detecting L.monocytogenes has a potential application.

Key concepts: Listeria monocytogenes, Loop-mediated isothermal amplification, Biology, DNA, Helicase, Microbiology, Polymerase chain reaction, Listeria

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