Reverse-transcriptase loop-mediated isothermal amplification as a rapid screening/monitoring tool for Salmonella enterica detection in liquid whole eggs

Journal of Food Science
Chayapa Techathuvanan, Doris Helen D'Souza

Abstract

Reverse-transcriptase loop-mediated isothermal amplification (RT-LAMP) is a novel molecular detection method that is specific, fast, and simple. It is based on reverse transcription followed by DNA amplification using the Bst DNA polymerase large fragment requiring one temperature and a simple waterbath, without the need for any expensive equipment. Detection is by turbidity or agarose gel electrophoresis. Our objective was to apply this LAMP-based technology to rapidly and sensitively detect Salmonella enterica serovar Enteritidis in liquid whole eggs (LWEs) within 1 d. LWE were inoculated with S. Enteritidis and stomached in tetrathionate broth (TTB), and spread-plated on Xylose lysine tergitol 4 agar either immediately or after 6, 12, or 16-h enrichment. RNA was extracted from 5-mL TTB and the RT-LAMP assay was carried out using invA primers. After 16 and 12-h enrichment, improved Salmonella detection up to 10⁰ to 10¹ and 10⁴ CFU/25 mL LWE, respectively was obtained. Without enrichment, Salmonella could be detected at 10⁷ CFU/25 mL; however, after 6-h enrichment a 1-log improvement to 10⁶ CFU/25 mL was obtained. This RT-LAMP assay appears to be suitable as a potential screening/monitoring tool for Salmonella enterica from LW...Continue Reading

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Citations

Jan 11, 2014·Journal of Animal Science and Biotechnology·Shijie FanGuiyun Xu
Feb 26, 2013·Journal of Agricultural and Food Chemistry·Felix FockeMarkus Fischer
Oct 12, 2013·Journal of Food Protection·Qianru YangBeilei Ge
May 23, 2018·Journal of Food Science·Chayapa Techathuvanan, Doris Helen D'Souza
Dec 7, 2016·Frontiers in Microbiology·Dexin ZengBaoguang Li

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