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Detection of Salmonella Typhi in Bile by Quantitative Real-Time PCR

In 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 import...

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Autores principales: Higginson, Ellen E., Nkeze, Joseph, Permala-Booth, Jasnehta, Kasumba, Irene N., Lagos, Rosanna, Hormazabal, Juan Carlos, Byrne, Alexander, Frankel, Gad, Levine, Myron M., Tennant, Sharon M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9241738/
https://www.ncbi.nlm.nih.gov/pubmed/35639002
http://dx.doi.org/10.1128/spectrum.00249-22
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author Higginson, Ellen E.
Nkeze, Joseph
Permala-Booth, Jasnehta
Kasumba, Irene N.
Lagos, Rosanna
Hormazabal, Juan Carlos
Byrne, Alexander
Frankel, Gad
Levine, Myron M.
Tennant, Sharon M.
author_facet Higginson, Ellen E.
Nkeze, Joseph
Permala-Booth, Jasnehta
Kasumba, Irene N.
Lagos, Rosanna
Hormazabal, Juan Carlos
Byrne, Alexander
Frankel, Gad
Levine, Myron M.
Tennant, Sharon M.
author_sort Higginson, Ellen E.
collection PubMed
description In 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 × 10(2) 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.
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spelling pubmed-92417382022-06-30 Detection of Salmonella Typhi in Bile by Quantitative Real-Time PCR Higginson, Ellen E. Nkeze, Joseph Permala-Booth, Jasnehta Kasumba, Irene N. Lagos, Rosanna Hormazabal, Juan Carlos Byrne, Alexander Frankel, Gad Levine, Myron M. Tennant, Sharon M. Microbiol Spectr Research Article In 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 × 10(2) 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. American Society for Microbiology 2022-05-31 /pmc/articles/PMC9241738/ /pubmed/35639002 http://dx.doi.org/10.1128/spectrum.00249-22 Text en Copyright © 2022 Higginson et al. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Article
Higginson, Ellen E.
Nkeze, Joseph
Permala-Booth, Jasnehta
Kasumba, Irene N.
Lagos, Rosanna
Hormazabal, Juan Carlos
Byrne, Alexander
Frankel, Gad
Levine, Myron M.
Tennant, Sharon M.
Detection of Salmonella Typhi in Bile by Quantitative Real-Time PCR
title Detection of Salmonella Typhi in Bile by Quantitative Real-Time PCR
title_full Detection of Salmonella Typhi in Bile by Quantitative Real-Time PCR
title_fullStr Detection of Salmonella Typhi in Bile by Quantitative Real-Time PCR
title_full_unstemmed Detection of Salmonella Typhi in Bile by Quantitative Real-Time PCR
title_short Detection of Salmonella Typhi in Bile by Quantitative Real-Time PCR
title_sort detection of salmonella typhi in bile by quantitative real-time pcr
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9241738/
https://www.ncbi.nlm.nih.gov/pubmed/35639002
http://dx.doi.org/10.1128/spectrum.00249-22
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