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Evaluation of PCR Approaches for Detection of Bartonella bacilliformis in Blood Samples

BACKGROUND: The lack of an effective diagnostic tool for Carrion’s disease leads to misdiagnosis, wrong treatments and perpetuation of asymptomatic carriers living in endemic areas. Conventional PCR approaches have been reported as a diagnostic technique. However, the detection limit of these techni...

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Autores principales: Gomes, Cláudia, Martinez-Puchol, Sandra, Pons, Maria J., Bazán, Jorge, Tinco, Carmen, del Valle, Juana, Ruiz, Joaquim
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4784731/
https://www.ncbi.nlm.nih.gov/pubmed/26959642
http://dx.doi.org/10.1371/journal.pntd.0004529
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author Gomes, Cláudia
Martinez-Puchol, Sandra
Pons, Maria J.
Bazán, Jorge
Tinco, Carmen
del Valle, Juana
Ruiz, Joaquim
author_facet Gomes, Cláudia
Martinez-Puchol, Sandra
Pons, Maria J.
Bazán, Jorge
Tinco, Carmen
del Valle, Juana
Ruiz, Joaquim
author_sort Gomes, Cláudia
collection PubMed
description BACKGROUND: The lack of an effective diagnostic tool for Carrion’s disease leads to misdiagnosis, wrong treatments and perpetuation of asymptomatic carriers living in endemic areas. Conventional PCR approaches have been reported as a diagnostic technique. However, the detection limit of these techniques is not clear as well as if its usefulness in low bacteriemia cases. The aim of this study was to evaluate the detection limit of 3 PCR approaches. METHODOLOGY/PRINCIPAL FINDINGS: We determined the detection limit of 3 different PCR approaches: Bartonella-specific 16S rRNA, fla and its genes. We also evaluated the viability of dry blood spots to be used as a sample transport system. Our results show that 16S rRNA PCR is the approach with a lowest detection limit, 5 CFU/μL, and thus, the best diagnostic PCR tool studied. Dry blood spots diminish the sensitivity of the assay. CONCLUSIONS/SIGNIFICANCE: From the tested PCRs, the 16S rRNA PCR-approach is the best to be used in the direct blood detection of acute cases of Carrion’s disease. However its use in samples from dry blood spots results in easier management of transport samples in rural areas, a slight decrease in the sensitivity was observed. The usefulness to detect by PCR the presence of low-bacteriemic or asymptomatic carriers is doubtful, showing the need to search for new more sensible techniques.
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spelling pubmed-47847312016-03-23 Evaluation of PCR Approaches for Detection of Bartonella bacilliformis in Blood Samples Gomes, Cláudia Martinez-Puchol, Sandra Pons, Maria J. Bazán, Jorge Tinco, Carmen del Valle, Juana Ruiz, Joaquim PLoS Negl Trop Dis Research Article BACKGROUND: The lack of an effective diagnostic tool for Carrion’s disease leads to misdiagnosis, wrong treatments and perpetuation of asymptomatic carriers living in endemic areas. Conventional PCR approaches have been reported as a diagnostic technique. However, the detection limit of these techniques is not clear as well as if its usefulness in low bacteriemia cases. The aim of this study was to evaluate the detection limit of 3 PCR approaches. METHODOLOGY/PRINCIPAL FINDINGS: We determined the detection limit of 3 different PCR approaches: Bartonella-specific 16S rRNA, fla and its genes. We also evaluated the viability of dry blood spots to be used as a sample transport system. Our results show that 16S rRNA PCR is the approach with a lowest detection limit, 5 CFU/μL, and thus, the best diagnostic PCR tool studied. Dry blood spots diminish the sensitivity of the assay. CONCLUSIONS/SIGNIFICANCE: From the tested PCRs, the 16S rRNA PCR-approach is the best to be used in the direct blood detection of acute cases of Carrion’s disease. However its use in samples from dry blood spots results in easier management of transport samples in rural areas, a slight decrease in the sensitivity was observed. The usefulness to detect by PCR the presence of low-bacteriemic or asymptomatic carriers is doubtful, showing the need to search for new more sensible techniques. Public Library of Science 2016-03-09 /pmc/articles/PMC4784731/ /pubmed/26959642 http://dx.doi.org/10.1371/journal.pntd.0004529 Text en © 2016 Gomes et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Gomes, Cláudia
Martinez-Puchol, Sandra
Pons, Maria J.
Bazán, Jorge
Tinco, Carmen
del Valle, Juana
Ruiz, Joaquim
Evaluation of PCR Approaches for Detection of Bartonella bacilliformis in Blood Samples
title Evaluation of PCR Approaches for Detection of Bartonella bacilliformis in Blood Samples
title_full Evaluation of PCR Approaches for Detection of Bartonella bacilliformis in Blood Samples
title_fullStr Evaluation of PCR Approaches for Detection of Bartonella bacilliformis in Blood Samples
title_full_unstemmed Evaluation of PCR Approaches for Detection of Bartonella bacilliformis in Blood Samples
title_short Evaluation of PCR Approaches for Detection of Bartonella bacilliformis in Blood Samples
title_sort evaluation of pcr approaches for detection of bartonella bacilliformis in blood samples
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4784731/
https://www.ncbi.nlm.nih.gov/pubmed/26959642
http://dx.doi.org/10.1371/journal.pntd.0004529
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