Study on the Rapid Detection of Salmonella by Loop Mediated Isothermal Amplification[Lamp Method].

Nanda Kumar, N.K. (2011) Study on the Rapid Detection of Salmonella by Loop Mediated Isothermal Amplification[Lamp Method]. [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:	In this study, an evaluation of the specificity and sensitivity of a Loop-mediated Isothermal amplification (LAMP) method for rapid detection of the food-borne pathogen Salmonella was carried out. A pair of outer primers and a pair of inner primers was specially designed for recognizing distinct sequences on the target invA gene. By the detection system, 241 bp target DNA was amplified and visualized as ladder like pattern of bands on agarose gel within 60 min under isothermal conditions at 65ºC. The detection limit of this LAMP assay was higher than that of PCR . Out of all the samples that was subjected to PCR analysis , only 23% gave positive results while the rest gave negative results . LAMP analysis was conducted on all the PCR positive samples , on some PCR negative samples and few other samples too . LAMP gave a much higher percentage of positive results , detecting very little quantity of 241 bp target DNA and amplifying it to a much better extend than PCR reaction. LAMP assay gave positive results for 80% of the samples tested and this was confirmed using 16s rRNA analysis . Spiking studies did not show much of a difference between PCR and LAMP , as both gave similar results . In addition, the LAMP method was carried out under isothermal condition, therefore no special apparatus was needed. A regular laboratory water bath or heating block is good enough. The LAMP method reported here demonstrated a potential and valuable means for detection of Salmonella , especially for its rapidity, simplicity and low cost.
Uncontrolled Keywords:	Loop-mediated Isothermal amplification, food-borne pathogen, Salmonella, detection
Subjects:	500 Natural Sciences and Mathematics > 04 Chemistry and Allied Sciences > 06 Chromatography 600 Technology > 01 Medical sciences > 07 Food borne disease
Divisions:	Fermentation Technology and Bioengineering
Depositing User:	Food Sci. & Technol. Information Services
Date Deposited:	08 Jul 2011 05:24
Last Modified:	28 Dec 2011 10:26
URI:	http://ir.cftri.res.in/id/eprint/10260

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