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dc.contributor.authorHigginson, Ellen E
dc.contributor.authorNkeze, Joseph
dc.contributor.authorPermala-Booth, Jasnehta
dc.contributor.authorKasumba, Irene N
dc.contributor.authorLagos, Rosanna
dc.contributor.authorHormazabal, Juan Carlos
dc.contributor.authorByrne, Alexander
dc.contributor.authorFrankel, Gad
dc.contributor.authorLevine, Myron M
dc.contributor.authorTennant, Sharon M
dc.date.accessioned2022-06-02T14:07:47Z
dc.date.available2022-06-02T14:07:47Z
dc.date.issued2022-05-31
dc.identifier.urihttp://hdl.handle.net/10713/19049
dc.description.abstractIn countries where the incidence of typhoid fever is high, fecal material from short-term carriers of Salmonella Typhi contaminates inadequately treated water supplies. As treated water supplies and improved sanitation become available, chronic (mainly gallbladder) carriers of S. Typhi become important. The objective of this study was to develop a method for detection of S. Typhi in bile by quantitative real-time PCR (qPCR) in patients undergoing cholecystectomy. We evaluated sensitivity and specificity of probesets that target oriC, viaB, fliC-d, STY0201, and stoD. We optimized DNA extraction from bile and compared the sensitivity of culture and our qPCR method to detect S. Typhi in bile samples containing various cephalosporins. With the use of an optimized DNA extraction technique, our limit of detection of S. Typhi in spiked human bile samples was 7.4 × 102 CFU/mL. We observed that S. Typhi could be detected by qPCR in samples containing cefazolin, cefotaxime, or ceftriaxone whereas culture could only detect Typhi in samples containing cefazolin but not cefotaxime or ceftriaxone. Our qPCR detection method for S. Typhi in bile should be preferred in areas where antibiotic usage is common. IMPORTANCE New Salmonella Typhi conjugate vaccines have been deployed, which will potentially lead to a fall in incidence rates of typhoid fever in endemic areas. Identification of chronic carriers of S. Typhi will be important as these individuals can be a potential source of transmission to susceptible persons. To address this public health concern, we have developed a novel method to detect S. Typhi in bile using real-time PCR. Our method can be used to identify carriers of S. Typhi among patients undergoing cholecystectomy (gallbladder removal surgery). The sensitivity of our molecular-based assay was superior to culture when performed in the presence of antibiotics commonly used during surgery. Our methodology will complement efforts to eliminate typhoid disease.en_US
dc.description.urihttps://doi.org/10.1128/spectrum.00249-22en_US
dc.language.isoenen_US
dc.publisherAmerican Society for Microbiologyen_US
dc.relation.ispartofMicrobiology Spectrumen_US
dc.subjectbileen_US
dc.subjectcarrieren_US
dc.subjectdetectionen_US
dc.subjectgall bladderen_US
dc.subjecttyphoiden_US
dc.titleDetection of Salmonella Typhi in Bile by Quantitative Real-Time PCR.en_US
dc.typeArticleen_US
dc.identifier.doi10.1128/spectrum.00249-22
dc.identifier.pmid35639002
dc.source.journaltitleMicrobiology spectrum
dc.source.beginpagee0024922
dc.source.endpage
dc.source.countryUnited States


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