Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Novel Heptaplex PCR-Based Diagnostics for Enteric Fever Caused by Typhoidal Salmonella Serovars and Its Applicability in Clinical Blood Culture

  • Hyun-Joong Kim (Department of Food Engineering, Mokpo National University) ;
  • Younsik Jung (Institute of Life Sciences and Resources and the Department of Food Science and Biotechnology, Kyung Hee University) ;
  • Mi-Ju Kim (Institute of Life Sciences and Resources and the Department of Food Science and Biotechnology, Kyung Hee University) ;
  • Hae-Yeong Kim (Institute of Life Sciences and Resources and the Department of Food Science and Biotechnology, Kyung Hee University)
  • Received : 2023.07.24
  • Accepted : 2023.08.14
  • Published : 2023.11.28
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Abstract

Enteric fever is caused by typhoidal Salmonella serovars (Typhi, Paratyphi A, Paratyphi B, and Paratyphi C). Owing to the importance of Salmonella serovars in clinics and public hygiene, reliable diagnostics for typhoidal serovars are crucial. This study aimed to develop a novel diagnostic tool for typhoidal Salmonella serovars and evaluate the use of human blood for clinically diagnosing enteric fever. Five genes were selected to produce specific PCR results against typhoidal Salmonella serovars based on the genes of Salmonella Typhi. Heptaplex PCR, including genetic markers of generic Salmonella, Salmonella enterica subsp. enterica, and typhoidal Salmonella serovars, was developed. Typhoidal Salmonella heptaplex PCR using genomic DNAs from 200 Salmonella strains (112 serovars) provided specifically amplified PCR products for each typhoidal Salmonella serovar. These results suggest that heptaplex PCR can sufficiently discriminate between typhoidal and non-typhoidal Salmonella serovars. Heptaplex PCR was applied to Salmonella-spiked blood cultures directly and provided diagnostic results after 12- or 13.5-h blood culture. Additionally, it demonstrated diagnostic performance with colonies recovered from a 6-h blood culture. This study provides a reliable DNA-based tool for diagnosing typhoidal Salmonella serovars that may be useful in clinical microbiology and epidemiology.

Keywords

Acknowledgement

This Research was supported by Research Funds of Mokpo National University in 2021. We are grateful for the clinical isolates of Salmonella Paratyphi A from the Asian Bacterial Bank (ABB) of the Asia Pacific Foundation for Infectious Diseases (APFID).

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