2014China Animal Husbandry & Veterinary MedicineRequires access

Establishment of Loop-mediated Isothermal Amplification Assay for Detection of Salmonella

LI Jia-ton

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Abstract

The assay was aimed to establish a rapid,sensitive and specific method of loop-mediated isothermal amplification(LAMP)for the detection of Salmonella.According to the highly conserved STN gene of Salmonellareported in GenBank, we designed a set of primers,and then followed by a series of LAMP optimization of reaction conditions,specificity test,sensitivity test,stability test and enzyme test.The sensitivity of LAMP and conventional PCR were compared.The results showed that the optimal reaction temperature of LAMP was 65 ℃,when only amplification of Salmonella,the minimum detectable concentration was 101 CFU/mL,which was more than conventional PCR detection by one order of magnitude.The results showed that LAMP method had the qualities of specificity,high sensitivity,short time-consuming and convenience for the detection of Salmonella,which was expected to develop into an effective method.

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What this paper is about

The assay was aimed to establish a rapid,sensitive and specific method of loop-mediated isothermal amplification(LAMP)for the detection of Salmonella.According to the highly conserved STN gene of Salmonellareported in GenBank, we designed a set of primers,and then followed by a series of LAMP optimization of reaction conditions,specificity test,sensitivity test,stability test and enzyme test.The sensitivity of LAMP and conventional PCR were compared.The results showed that the optimal reaction temperature of LAMP was 65 ℃,when only amplification of Salmonella,the minimum detectable concentration was 101 CFU/mL,which was more than conventional PCR detection by one order of magnitude.The results showed that LAMP method had the qualities of specificity,high sensitivity,short time-consuming and convenience for the detection of Salmonella,which was expected to develop into an effective method.

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Available abstract

The assay was aimed to establish a rapid,sensitive and specific method of loop-mediated isothermal amplification(LAMP)for the detection of Salmonella.According to the highly conserved STN gene of Salmonellareported in GenBank, we designed a set of primers,and then followed by a series of LAMP optimization of reaction conditions,specificity test,sensitivity test,stability test and enzyme test.The sensitivity of LAMP and conventional PCR were compared.The results showed that the optimal reaction temperature of LAMP was 65 ℃,when only amplification of Salmonella,the minimum detectable concentration was 101 CFU/mL,which was more than conventional PCR detection by one order of magnitude.The results showed that LAMP method had the qualities of specificity,high sensitivity,short time-consuming and convenience for the detection of Salmonella,which was expected to develop into an effective method.

Key concepts: Loop-mediated isothermal amplification, Salmonella, Sensitivity (control systems), GenBank, Detection limit, Molecular biology, Isothermal process, Biology

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