Publication:
Molecular detection of Salmonella enterica serovar Typhi by Vi-qPCR

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dc.contributor.authorShakrin, NNSMen_US
dc.contributor.authorAdnan, SNAen_US
dc.contributor.authorWahab, AHAen_US
dc.contributor.authorRamasamy, RPen_US
dc.contributor.authorYussof, WNWen_US
dc.contributor.authorNoordin, Nen_US
dc.contributor.authorVerashahib, Ken_US
dc.contributor.authorJahis, Ren_US
dc.date.accessioned2024-05-29T02:49:37Z
dc.date.available2024-05-29T02:49:37Z
dc.date.issued2018
dc.description.abstractAims: To develop a real-time polymerase chain reaction system Vi-qPCR in the detection of Salmonella enterica serovar Typhi (S. Typhi), targeting the vexC gene encoding for Vi antigen (capsular polysaccharide antigen) and to evaluate its sensitivity and specificity performance using pure cultures of S. Typhi and other enteric pathogens. Methodology and results: Microbiological, biochemical and serotyping tests were conducted to determine the phenotypic characteristics of S. Typhi and other enteric pathogens in our collection. Primers were designed using Primer3 software and their in-silico specificity were analysed using Basic Local Alignment System Tool (BLAST). Optimisation of PCR annealing temperature was done prior to assessment of sensitivity and specificity performance against artificial serially diluted seeded stools. The primers were found to be 100% specific in the detection of S. Typhi towards 32 tested clinical strains. Verification of gene amplification by comparing the nucleotide sequences against reference genes in the GenBank database revealed high specificity to S. Typhi. Statistical analysis indicates that this method results in 100% sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV). Moreover, Vi-qPCR allows the detection of S. Typhi as low as 13 +/- 1.4 CFU/g of stool sample. Conclusion, significance and impact of study: A rapid and sensitive method for detection of Salmonella enterica serovar Typhi (S. Typhi) is desired as a diagnostic tool to improve typhoid management. The Vi-qPCR represent a promising non-invasive diagnostic tool for medical microbiology laboratories as a method for the detection of S. Typhi in both pure culture and stool specimens especially in chronic asymptomatic carriers where shedding of S. Typhi is intermittent and sometimes occurs in low level.en_US
dc.identifier.doi10.21161/mjm.1461806
dc.identifier.epage489
dc.identifier.isbn2231-7538
dc.identifier.issn1823-8262
dc.identifier.issue6
dc.identifier.scopusWOS:000456110600006
dc.identifier.spage483
dc.identifier.urihttps://oarep.usim.edu.my/handle/123456789/10889
dc.identifier.volume14
dc.languageEnglish
dc.language.isoen_USen_US
dc.publisherMalaysian Soc Microbiologyen_US
dc.relation.ispartofMalaysian Journal Of Microbiologyen_US
dc.sourceWeb Of Science (ISI)
dc.subjectPCRen_US
dc.subjectdiagnosticen_US
dc.subjectpathogenen_US
dc.titleMolecular detection of Salmonella enterica serovar Typhi by Vi-qPCRen_US
dc.typeArticleen_US
dspace.entity.typePublication

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