Development and evaluation of alternative methods to identify the three most common serotypes of salmonella enterica causing clinical infections in Kazakhstan
Barmak S.M. Sinyavskiy Y.A. Berdygaliev A.B. Sharmanov T.S. Savitskaya I.S. Sultankulova G.T. Zholdybayeva E.V.
November 2021MDPI
Microorganisms
2021#9Issue 11
In this study, we aimed to compare the performance of conventional PCR and real-time PCR assays as screening methods for identification of three frequent, clinically significant Salmonella serovars in Kazakhstan. We determined the diagnostic efficacy of three molecular methods for detection of S. enterica subsp. enterica and typing S. Typhimurium, S. Enteritidis, and S. Virchow. A total of 137 clinical samples and 883 food samples were obtained in Almaty in 2018–2019. All tests showed high analytical specificity for detecting S. enterica and its corresponding serovariants (100%). The sensitivity of real-time PCR for each of the tested targets was 1–10 microbial cells and in conventional PCR 10–100 microbial cells. The trials with conventional PCR and real-time PCR had a diagnostic efficacy (DE) of 100% and 99.71%, respectively. The DE of real-time PCR and conventional PCR for detecting S. Enteritidis and S. Typhimurium was 99.90%, while the DE of conventional PCR and real-time PCR for detecting S. Virchow was 99.31% and 99.80%, respectively. The RAPD-PCR analysis of the genomic DNA of Salmonella enterica showed the genetic kinship of S. Enteritidis isolates, and the genetic heterogeneity of S. Typhimurium and S. Virchow isolates. Thus, the developed methods can be considered as alternatives to classical serotyping using antisera.
PCR , RAPD-PCR , Real-time PCR , S. Enteritidis , S. Typhimurium , S. Virchow , Salmonella enterica
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Faculty of Biology and Biotechnology, Al-Farabi Kazakh National University, Almaty, 050040, Kazakhstan
Biotechnology and Biologically Active Substances Laboratory, Kazakh Academy of Nutrition, Almaty, 050008, Kazakhstan
Department of Pediatric Surgery, Asfendiyarov Kazakh National Medical University, Almaty, 050012, Kazakhstan
National Center for Biotechnology, National Scientific Shared Laboratory of Biotechnology, Nur-Sultan, 010000, Kazakhstan
Faculty of Biology and Biotechnology
Biotechnology and Biologically Active Substances Laboratory
Department of Pediatric Surgery
National Center for Biotechnology
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