Development of a novel multiplex PCR for the detection and differentiation of Salmonella enterica serovars Typhi and Paratyphi A

In this study, we developed a multiplex polymerase chain reaction (mPCR) assay for pan- Salmonella detection as well as for specific detection of serovars Typhi and Paratyphi A. The assay detects members of the Salmonella genus by amplifying the outer membrane protein C ( ompC). The presence of eith...

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Bibliographic Details
Published in:Research in microbiology 2010-05, Vol.161 (4), p.243-248
Main Authors: Yin Ngan, Grace Jie, Ng, Li Mei, Lin, Raymond T.P., Teo, Jeanette W.P.
Format: Article
Language:English
Subjects:
Bacterial Proteins - genetics
Bacterial Typing Techniques - methods
Bacteriology
Biological and medical sciences
Colony-forming cells
Comparative genomics
Enteric fever
Fundamental and applied biological sciences. Psychology
Humans
Microbiology
Miscellaneous
Multiplex polymerase chain reaction
Paratyphis
Phylogeny
Polymerase Chain Reaction - methods
Porins - genetics
Salmonella enterica serovar Paratyphi A
Salmonella enterica serovar Typhi
Salmonella Infections - microbiology
Salmonella paratyphi A - classification
Salmonella paratyphi A - genetics
Salmonella paratyphi A - isolation & purification
Salmonella typhi - classification
Salmonella typhi - genetics
Salmonella typhi - isolation & purification
Sequence Analysis, DNA
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Summary:In this study, we developed a multiplex polymerase chain reaction (mPCR) assay for pan- Salmonella detection as well as for specific detection of serovars Typhi and Paratyphi A. The assay detects members of the Salmonella genus by amplifying the outer membrane protein C ( ompC). The presence of either Salmonella serovar Typhi or Paratyphi A is indicated by amplification of the putative regulatory protein gene STY4220, which is common to both serovars. Further differentiation of the serovars was achieved by targeting the intergenic region (SSPAI) between SSPA1723a and SSPA1724 in serovar Paratyphi A, and stgA, a fimbrial subunit protein, in serovar Typhi. mPCR was evaluated using 124 clinical and reference Salmonella serovars. S. enterica serovars Typhi and Paratyphi A were detected at 100% specificity and sensitivity. Each PCR reaction detected approximately 1 pg of Salmonella genomic DNA. Sensitivity of the PCR when tested on 8-h-enriched spiked blood samples of serovars Typhi and Paratyphi A was estimated at 4.5 × 10 4–5.5 × 10 4 CFU/ml, with similar detection levels observed for spiked fecal samples. This mPCR could prove to be a useful diagnostic tool for the detection and differentiation of serovars Typhi and Paratyphi A.
ISSN:0923-2508
1769-7123
DOI:10.1016/j.resmic.2010.03.005