DETECTION OF Salmonella spp. IN FOOD BY LAMP (LOOP-MEDIATED ISOTHERMAL AMPLIFICATION) METHOD

  • Nguyễn Phạm Trúc Phương, Đoàn Thị Quỳnh Hương, Nguyễn Văn Lượng
Keywords: Loop Mediated Isothermal Amplification; invA gene on Salmonella; Salmonella enteritidis; Food; SYBR® Green I nucleic acid gel stain

Abstract

Method for early and rapid detection of foodborne pathogens caused by Salmonella bacteria will help control contaminated food in the market. Therefore, in this study, a procedure for detecting Salmonella in foods by LAMP technique has been developed to quickly and accurately detect strains of Salmonella spp. based on the invA2 primer set specifically for the invA gene region of Salmonella. The products were visualised directly by colour changes of the reaction. Positive results were indicated by green fluorescence and negative by orange colour. The test was further evaluated for specificity, sensitivity. The optimized LAMP protocol include concentrations of primers F3B3/FIPBIP 200Nm/1600nM (1:8) and Betaine 0,8M, 60 minutes heating at 63oC, the specificity of LAMP protocol for the detection of Salmonella spp. was 100% while sensitivity was 5pg/reaction (on DNA template), 4.1x101CFU/ml (on bacterial strain). The results were compared with those obtained by realtime PCR.

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Published
2022-01-28
Section
AGRICULTURE – FORESTRY – MEDICINE & PHARMACY