[Rapid identification of 15 common S. enterica Salmonella serovars by multiplex real-time PCR].

OBJECTIVE

Multiplex real-time PCR for the identification of 15 Salmonella serovars was developed.

METHODS

Through the Salmonella genome comparison, 12 membrane proteins STM4497 gene can be used to identify 15 Salmonella serovars, and these 12 genes were respectively listed as A-L genes. Then primers were designed according to A-L gene conserved sequences, and then multiplex real-time PCR was established assessed with the evaluation of the limit detection, sensitivity, specificity, and repeatability. The 206 Salmonella strains were identified using multiplex real-time PCR with the comparison of the serum slide agglutination assay.

RESULTS

The limit detection of multiplex PCR ranged from 1. 1×10~(-3)-1. 2×10~(-3) ng/μL. The target genes were 100% specificity, and the relative standard deviation was lower than 2. 97%. Compared with the serum slide agglutination assay, Kappa ranged 0. 92-1. 00.

CONCLUSION

The multiplex real-time PCR can be used to identify 15 Salmonella serovars, which is rapid, accurate and specific.