Evaluation of primer detection capabilities of fimC Salmonella typhi using real time PCR for rapid detection of bacteria causes of food poisoning

Nurjayadi, Muktiningsih; Islami, N.; Pertiwi, Yuniariana; Saamia, V.; Wirana, I M

doi:10.1088/1757-899x/434/1/012097

Cited by 3 publications

(1 citation statement)

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“…Various molecular methods for identi cation of typhoidal/nontyphoidal Salmonella had previously developed (18, 29) but most of them focused on just one or two of the EF pathogens. Only one single target gene chosen often means much lower speci city, like mC gene for S. typhi, which still requires improvement in its speci city (30). In addition, discriminating S.Paratyphi B from S.Paratyphi B variant Java is important for the food chain industry, because the pathogenicity of these two is totally different.…”

Section: Discussionmentioning

confidence: 99%

Rapid and Specific Differentiation of Salmonella enterica serotypes Typhi and Paratyphi by Multicolor Melting Curve Analysis

Jiang,

Cai

et al. 2024

Preprint

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Rapid and accurate identification of Salmonella enterica serotypes Typhi and Paratyphi (A, B and C), the causal agents of enteric fever, is critical for timely treatment, case management and evaluation of health policies in low and middle-income countries where the disease still remains a serious public health problem. The present study describes the development of a multiplex assay (EFMAtyping) for simultaneous identification of pathogens causing typhoid and paratyphoid fever in a single reaction by the MeltArray approach, which could be finished within 2.5 h. Seven specific genes were chosen for differentiation of typhoidal and nontyphoidal Salmonella. All gene targets were able to be detected by the EFMAtyping assay, with expected Tm values and without cross-reactivity to other relevant Salmonella serovars. The limit of detection (LOD) for all gene targets was 50 copies per reaction. The LOD reached 102-103 CFU/ml for each pathogen in simulated clinical samples. The largest standard deviation value for mean Tm was below 0.5°C. This newly developed EFMAtyping assay was further evaluated by testing 551 clinical Salmonella isolates, corroborated in parallel by the traditional Salmonella identification workflow, and serotype prediction was enabled by whole-genome sequencing. Compared to the traditional method, our results exhibited 100% of specificity and greater than 96% of sensitivity with a kappa correlation ranging from 0.96 to 1.00. Thus, the EFMAtyping assay provides a rapid, high throughput, and promising tool for public health laboratories to monitor typhoid and paratyphoid fever.

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Section: Discussionmentioning

confidence: 99%