Rapid, Sensitive, Specific, and Visual Detection of Salmonella in Retail Meat with Loop-Mediated Isothermal Amplification, Targeting the invA Gene

J Food Prot. 2022 Jan 1;85(1):6-12. doi: 10.4315/JFP-21-186.

Abstract

Abstract: Salmonella is one of the major pathogenic bacteria causing foodborne diseases. The rapid detection of Salmonella in food is of great significance to food safety. In this study, the loop-mediated isothermal amplification (LAMP) method was developed, and primers were designed targeting the invA gene of Salmonella. Standard samples of recombinant invA-plasmid and 100 retail meat samples were tested by LAMP and compared with the results tested by conventional PCR and the routine Chinese National Food Safety Standard-Microbiological Examination of Food-Examination of Salmonella, respectively. The results showed that Salmonella strains of eight different serotypes were amplified successfully by the developed LAMP assay, and it was 1,000-fold more sensitive than conventional PCR, with the analytical sensitivity of 8 × 102 copies per μL of the standard sample of invA-plasmid. The results were visualized directly by adding calcein and MnCl2 in the LAMP reaction tube, and the positively amplified products turned green after an incubation of 2 min. In parallel detection, the positive rate of Salmonella by the LAMP assay was highly correlated with the routine Chinese national standard method. The results of the study demonstrated that the developed LAMP assay is a simple, rapid, strongly specific, highly sensitive, and visual detection method for Salmonella.

Keywords: Salmonella; invA Gene; Loop-mediated isothermal amplification; Rapid detection; Retail meat; Visual detection.

MeSH terms

  • Meat / microbiology
  • Molecular Diagnostic Techniques
  • Nucleic Acid Amplification Techniques* / methods
  • Salmonella* / genetics
  • Sensitivity and Specificity

Supplementary concepts

  • LAMP assay