2011Food Science and Technology InternationalRequires access

Development of a loop-mediated isothermal amplification forthe detection of Yersinia enterocolitica

Wei Zhang

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Abstract

A loop-mediated isothermal amplification(LAMP) technology with two loop primers was developed for rapidly detecting Yersinia enterocolitica in chicken. Sequences of Ail gene of Y.e (AY004311.1) were used as target sequences, to design outer primers, inner primers. We judged the results of detection, through visible to the naked eye of the white precipitate. The result indicates: we extracted DNA using the method of Chemical reagent. The detection limit of pure bacterial culture was 6.3 cfu/mL and the detection limit of artificially contaminated chicken was 340 cfu/g with improved LAMP detection for two hours. In contrast, the detection limit of pure bacterial culture was 630 cfu/mL and the detection limit of artificially contaminated chicken was 3.4×104 cfu/g with PCR detection for three hours. Therefore, LAMP has the potential to replace PCR because of its simplicity, rapidity, specificity, no special equipment, and cost-effectiveness. It is very suitable for the majority of the our local laboratory on the application. For the rapid detection of foodborne pathogens build a technology platform.

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What this paper is about

A loop-mediated isothermal amplification(LAMP) technology with two loop primers was developed for rapidly detecting Yersinia enterocolitica in chicken. Sequences of Ail gene of Y.e (AY004311.1) were used as target sequences, to design outer primers, inner primers. We judged the results of detection, through visible to the naked eye of the white precipitate. The result indicates: we extracted DNA using the method of Chemical reagent. The detection limit of pure bacterial culture was 6.3 cfu/mL and the detection limit of artificially contaminated chicken was 340 cfu/g with improved LAMP detection for two hours. In contrast, the detection limit of pure bacterial culture was 630 cfu/mL and the detection limit of artificially contaminated chicken was 3.4×104 cfu/g with PCR detection for three hours. Therefore, LAMP has the potential to replace PCR because of its simplicity, rapidity, specificity, no special equipment, and cost-effectiveness. It is very suitable for the majority of the our local laboratory on the application. For the rapid detection of foodborne pathogens build a technology platform.

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Available abstract

A loop-mediated isothermal amplification(LAMP) technology with two loop primers was developed for rapidly detecting Yersinia enterocolitica in chicken. Sequences of Ail gene of Y.e (AY004311.1) were used as target sequences, to design outer primers, inner primers. We judged the results of detection, through visible to the naked eye of the white precipitate. The result indicates: we extracted DNA using the method of Chemical reagent. The detection limit of pure bacterial culture was 6.3 cfu/mL and the detection limit of artificially contaminated chicken was 340 cfu/g with improved LAMP detection for two hours. In contrast, the detection limit of pure bacterial culture was 630 cfu/mL and the detection limit of artificially contaminated chicken was 3.4×104 cfu/g with PCR detection for three hours. Therefore, LAMP has the potential to replace PCR because of its simplicity, rapidity, specificity, no special equipment, and cost-effectiveness. It is very suitable for the majority of the our local laboratory on the application. For the rapid detection of foodborne pathogens build a technology platform.

Key concepts: Loop-mediated isothermal amplification, Yersinia enterocolitica, Detection limit, Contamination, Molecular biology, Reagent, Biology, Microbiology

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