Study on the rapid detection of V.cholerae by loop-mediated isothermal amplification method from food and water samples.

Himanshu, Mahawar (2011) Study on the rapid detection of V.cholerae by loop-mediated isothermal amplification method from food and water samples. [Student Project Report]

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Abstract

This Dissertation / Report is the outcome of investigation carried out by the creator(s) / author(s) at the department/division of Central Food Technological Research Institute (CFTRI), Mysore mentioned below in this page.

Item Type:	Student Project Report
Additional Information:	The purpose of this study was to establish a loop-mediated isothermal amplification (LAMP) method for the rapid and sensitive detection of V.cholerae from food and water samples. Detection of ctxA producing V.cholerae using conventional techniques like culture, biochemical and immunological based assays are time consuming and laborious. Thus we developed a highly specific loop-mediated isothermal amplification (LAMP) assay for the sensitive and rapid detection of cholera toxin producing V.cholerae. Direct lysis method was used to extract DNA from food samples by using 1% triton-X 100. The LAMP reaction was carried out at 65oC for 60 min. Unlike PCR, LAMP reaction was not affected by the inhibitors of the food components. The amplified products were detected by visual inspection using SYBR Green I dye. The assay provided markedly more sensitive and rapid detection of ctxA producing V.cholerae strains than conventional biochemical and PCR assays. The detection sensitivity of LAMP, PCR and biochemical tests were found to be 82%, 19% and 20%, respectively. The LAMP assay is a sensitive, rapid and simple tool for the detection of V.cholerae producing cholera toxin. This technique provides an important detecting tool for the detection of V.cholerae infection both in the laboratory and field. This technique can be recommended as an applied protocol for food safety analysis, health management programme and disease surveillance. This method provides us with an alternate powerful tool in food safety and diagnosis of infectious disease.
Uncontrolled Keywords:	loop-mediated isothermal amplification, V.cholerae, food, water samples
Subjects:	600 Technology > 08 Food technology > 10 Food Microorganisms
Divisions:	Fermentation Technology and Bioengineering
Depositing User:	Food Sci. & Technol. Information Services
Date Deposited:	07 Jul 2011 09:27
Last Modified:	28 Dec 2011 10:25
URI:	http://ir.cftri.res.in/id/eprint/10237

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