Teh, Cindy Shuan Ju and Lau, Min Yi and Chong, Chun Wie and Ngoi, Soo Tein and Chua, Kek Heng and Lee, Way Seah and Thong, Kwai Lin (2021) One-step differential detection of Salmonella enterica serovar Typhi, serovar Paratyphi A and other Salmonella spp. by using a quadruplex real-time PCR assay. Journal of Microbiological Methods, 183. ISSN 0167-7012, DOI https://doi.org/10.1016/j.mimet.2021.106184.
Full text not available from this repository.Abstract
Diseases caused by typhoidal and non-typhoidal Salmonella remain a considerable threat to both developed and developing countries. Based on the clinical symptoms and serological tests, it is sometimes difficult to differentiate the Salmonella enterica serovar Paratyphi A (S. enterica serovar Paratyphi A) from semvar Typhi (S. enterica serovar Typhi). In this study, we developed a quadruplex real-time polymerase chain reaction (PCR) assay with an internal amplification control (IAC), to simultaneously differentiate S. enterica serovar Paratyphi A from semvar Typhi and to detect other Salmonella serovars which cause salmonellosis in humans. This assay was evaluated on 155 salmonellae and non-salmonellae strains and demonstrated 100% specificity in species differentiation. Inclusion of an IAC did not affect the efficiency of the assay. Further evaluation using a blind test on spiked stool, blood and food specimens showed that the detection limit was at 10(3) -10(4) CFU/mL (or g) and a high PCR efficiency with different targets (R-2 > 0.99), except for S. enterica semvar Paratyphi A in blood. This assay has been applied to clinical specimens to detect the causative agents of gastrointestinal infections and has successfully identified 6 salmonellosis patients from the 50 diarrhoea patients. The quadruplex real-time PCR developed in this study could enhance the detection and differentiation of salmonellae. This assay could be applied to stools, blood and food based on the notable performance in the simulation tests and field evaluation.
| Item Type: | Article |
|---|---|
| Funders: | International Fund received by the Principal Investigator [Grant No: IF004-2020]. |
| Uncontrolled Keywords: | S. enterica serovar Typhi; S. enterica serovar Paratyphi A; Real-time PCR; Salmonellae detection |
| Subjects: | Q Science > QR Microbiology |
| Divisions: | Faculty of Medicine Deputy Vice Chancellor (Research & Innovation) Office > Nanotechnology & Catalysis Research Centre |
| Depositing User: | Ms Zaharah Ramly |
| Date Deposited: | 21 Apr 2022 06:08 |
| Last Modified: | 21 Apr 2022 06:08 |
| URI: | http://eprints.um.edu.my/id/eprint/28827 |
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