Evaluation of three PCR schemes for detection of Bartonella bacilliformis in blood samples: sensitivity, specificity and applicability

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Revisión por pares
Detalles Bibliográficos
Autores: Gomes, C.S.P., Silva, W., Tinco, C., Martinez Puchol, S., Pons, Maria J, Bazan, Jorge, Del Valle Mendoza, Juana, Ruiz, J., Universidad Peruana de Ciencias Aplicadas (UPC)
Formato: objeto de conferencia
Fecha de Publicación:2015
Institución:Universidad Peruana de Ciencias Aplicadas
Repositorio:UPC-Institucional
Lenguaje:inglés
OAI Identifier:oai:repositorioacademico.upc.edu.pe:10757/347086
Enlace del recurso:http://hdl.handle.net/10757/347086
Nivel de acceso:acceso abierto
Materia:https://purl.org/pe-repo/ocde/ford#3.00.00
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dc.title.es_PE.fl_str_mv Evaluation of three PCR schemes for detection of Bartonella bacilliformis in blood samples: sensitivity, specificity and applicability
title Evaluation of three PCR schemes for detection of Bartonella bacilliformis in blood samples: sensitivity, specificity and applicability
spellingShingle Evaluation of three PCR schemes for detection of Bartonella bacilliformis in blood samples: sensitivity, specificity and applicability
Gomes, C.S.P.
https://purl.org/pe-repo/ocde/ford#3.00.00
title_short Evaluation of three PCR schemes for detection of Bartonella bacilliformis in blood samples: sensitivity, specificity and applicability
title_full Evaluation of three PCR schemes for detection of Bartonella bacilliformis in blood samples: sensitivity, specificity and applicability
title_fullStr Evaluation of three PCR schemes for detection of Bartonella bacilliformis in blood samples: sensitivity, specificity and applicability
title_full_unstemmed Evaluation of three PCR schemes for detection of Bartonella bacilliformis in blood samples: sensitivity, specificity and applicability
title_sort Evaluation of three PCR schemes for detection of Bartonella bacilliformis in blood samples: sensitivity, specificity and applicability
dc.creator.es_PE.fl_str_mv Universidad Peruana de Ciencias Aplicadas (UPC)
author Gomes, C.S.P.
author_facet Gomes, C.S.P.
Silva, W.
Tinco, C.
Martinez Puchol, S.
Pons, Maria J
Bazan, Jorge
Del Valle Mendoza, Juana
Ruiz, J.
Universidad Peruana de Ciencias Aplicadas (UPC)
author_role author
author2 Silva, W.
Tinco, C.
Martinez Puchol, S.
Pons, Maria J
Bazan, Jorge
Del Valle Mendoza, Juana
Ruiz, J.
Universidad Peruana de Ciencias Aplicadas (UPC)
author2_role author
author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Gomes, C.S.P.
Silva, W.
Tinco, C.
Martinez Puchol, S.
Pons, Maria J
Bazan, Jorge
Del Valle Mendoza, Juana
Ruiz, J.
dc.subject.ocde.es_PE.fl_str_mv https://purl.org/pe-repo/ocde/ford#3.00.00
topic https://purl.org/pe-repo/ocde/ford#3.00.00
description Revisión por pares
publishDate 2015
dc.date.accessioned.es_PE.fl_str_mv 2015-03-24T19:41:23Z
dc.date.available.es_PE.fl_str_mv 2015-03-24T19:41:23Z
dc.date.issued.fl_str_mv 2015-03-24
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dc.identifier.issn.es_PE.fl_str_mv 1201-9712
dc.identifier.uri.es_PE.fl_str_mv http://hdl.handle.net/10757/347086
identifier_str_mv 1201-9712
url http://hdl.handle.net/10757/347086
dc.language.iso.es_PE.fl_str_mv eng
language eng
dc.rights.es_PE.fl_str_mv info:eu-repo/semantics/openAccess
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dc.format.es_PE.fl_str_mv application/pdf
dc.publisher.es_PE.fl_str_mv Elsevier B.V.
dc.source.es_PE.fl_str_mv Universidad Peruana de Ciencias Aplicadas (UPC)
Repositorio Académico - UPC
dc.source.none.fl_str_mv reponame:UPC-Institucional
instname:Universidad Peruana de Ciencias Aplicadas
instacron:UPC
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reponame_str UPC-Institucional
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spelling 515a5adbde9111a13f6a7a625ec257fd-192ab769a02c4a073866d9e46a4da86f6-1937c649056ba843bfd8afeec8df370cf-12ea0c51cd9ab1fc6165c9fb07a210cad-1e31e21e8cc904fc5feb3e9f70bccc057-1c9f564b5fb2681f567c5d401b13962ac-1f92b24b86b6890d5ff7acafea7fac632-1ea4494b3c2da7e9ff48f961207fed373-1Gomes, C.S.P.Silva, W.Tinco, C.Martinez Puchol, S.Pons, Maria JBazan, JorgeDel Valle Mendoza, JuanaRuiz, J.Universidad Peruana de Ciencias Aplicadas (UPC)2015-03-24T19:41:23Z2015-03-24T19:41:23Z2015-03-241201-9712http://hdl.handle.net/10757/347086Revisión por pares16th International Congress on Infectious Diseases (ICID), 2014. 2-5 de Abril 2014. Cape Town, South AfricaBackground: Bartonella bacilliformis is the etiological agent of Carrion’s disease, a neglected illness with a febrile lethal stage and a warty benign phase, being the human the only known reservoir. The diagnostic by microscopy in endemic areas is several times erroneous. Furthermore, the culture of this bacterium is time-consuming, being the diagnostic by PCR the easiest way to perform a correct diagnostic. The objective of this study was to evaluate the detection limit of three PCR schemes, designed to detect B.bacilliformis, both in blood and filter papers to test their potential use for transferring samples from endemic areas to reference centers. Moreover, the specificity was also observed as well as the applicability of the technique with clinical samples from different stages of the disease. Methods & Materials: Fragments of 16SrRNA and fla genes were amplified as well as the variable-intergenic region (its). The detection limit was determined by bacterial quantification with flow cytometry and performing dilutions (106cfu/ml-10cfu/ml) both in blood and filter papers. DNA was extracted and PCRs were performed. Specificity was tested by processing other bacteraemia microorganisms. Clinical samples, 12 from febrile patients, 13 from warty and 71 from healthy asymptomatic individuals living in endemic area(Mandinga-Cajamarca) were also processed. Results: The 16SrRNA PCR scheme showed the lower detection limit (5 cfu from blood and filter paper) being the PCR scheme chosen to be tested in clinical samples. All febrile patients’ samples were positive, whereas in warty individuals only 3(23%) faint bands were obtained. No amplification was obtained in samples from healthy people. Fainter bands were always obtained when PCRs were made of filter papers. All PCRs were specific for B.bacilliformis. Conclusion: The 16SrRNA PCR seems to be the best technique to detect feverish patients. However, the applicability to identify asymptomatic carriers was undetermined. Filter paper may be an alternative for easy transportation of samples but is need to consider the decreasing sensitivity of the results. It is critical to develop rapid, sensitive and specific technique capable of being applied in endemic rural areas, to avoid misdiagnosis and facilitate the detection of asymptomatic carriers that will allow progress towards the eradication of this disease.application/pdfengElsevier B.V.info:eu-repo/semantics/openAccessUniversidad Peruana de Ciencias Aplicadas (UPC)Repositorio Académico - UPCreponame:UPC-Institucionalinstname:Universidad Peruana de Ciencias Aplicadasinstacron:UPCEvaluation of three PCR schemes for detection of Bartonella bacilliformis in blood samples: sensitivity, specificity and applicabilityinfo:eu-repo/semantics/conferenceObjecthttps://purl.org/pe-repo/ocde/ford#3.00.002018-06-23T09:14:35ZBackground: Bartonella bacilliformis is the etiological agent of Carrion’s disease, a neglected illness with a febrile lethal stage and a warty benign phase, being the human the only known reservoir. The diagnostic by microscopy in endemic areas is several times erroneous. Furthermore, the culture of this bacterium is time-consuming, being the diagnostic by PCR the easiest way to perform a correct diagnostic. The objective of this study was to evaluate the detection limit of three PCR schemes, designed to detect B.bacilliformis, both in blood and filter papers to test their potential use for transferring samples from endemic areas to reference centers. Moreover, the specificity was also observed as well as the applicability of the technique with clinical samples from different stages of the disease. Methods & Materials: Fragments of 16SrRNA and fla genes were amplified as well as the variable-intergenic region (its). The detection limit was determined by bacterial quantification with flow cytometry and performing dilutions (106cfu/ml-10cfu/ml) both in blood and filter papers. DNA was extracted and PCRs were performed. Specificity was tested by processing other bacteraemia microorganisms. Clinical samples, 12 from febrile patients, 13 from warty and 71 from healthy asymptomatic individuals living in endemic area(Mandinga-Cajamarca) were also processed. Results: The 16SrRNA PCR scheme showed the lower detection limit (5 cfu from blood and filter paper) being the PCR scheme chosen to be tested in clinical samples. All febrile patients’ samples were positive, whereas in warty individuals only 3(23%) faint bands were obtained. No amplification was obtained in samples from healthy people. Fainter bands were always obtained when PCRs were made of filter papers. All PCRs were specific for B.bacilliformis. Conclusion: The 16SrRNA PCR seems to be the best technique to detect feverish patients. However, the applicability to identify asymptomatic carriers was undetermined. Filter paper may be an alternative for easy transportation of samples but is need to consider the decreasing sensitivity of the results. 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Evaluation of three PCR schemes for detection of Bartonella bacilliformis in blood samples: sensitivity, specificity and applicability
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