Aplicación de ensayos de PCR en tiempo real para el diagnóstico de la histoplasmosis utilizando tres dianas moleculares en tejidos humanos de FFPE y sangre.

Luisa F. López, Oliver Keatinge Clay, Diego H. Cáceres, Beatriz L. Gómez

Resultado de la investigación: Tipos de Contribuciónes en ConferenciaResumen

Resumen

Objective: Histoplasmosis is a fungal infection that causes significantmorbidity andmortality in persons living withHIV/AIDS, especially in countries with limited resources. Currently used diagnostic tests rely on culture and serology, lack sensitivity and often require weeks to obtain results causing significant diagnosis delays; molecular assays are not commercially available. Methods: we aimed to apply quantitative real-time PCR (qPCR) targeting three protein-coding genes of Histoplasma capsulatum (100-kDa, H and M antigens) for detection of H. capsulatum infection in formalin-fixed paraffin-embedded (FFPE) and whole blood (WB) samples from patients with proven histoplasmosis. Results: For FFPE samples, the sensitivity of 100-kDa, H and M qPCR assays were 93.9%, 91% and 57%, respectively; however, the same qPCR assays showed only 23%, 19% and 11.5% of sensitivity for 100-kDa, H y M qPCR assays when used with the WB samples. The specificity of qPCR was determined by testing samples from patients with other clinical infections and healthy controls and was 93%-100% depending upon the assay and the specimen type. Conclusion: we applied three qPCR assays for detecting H. capsulatum DNA in human samples, and demonstrated that the molecular protocols based on amplification of 100-kDa and H antigen can be successfully used for diagnosing this mycosis when using FFPE samples; however, we do not recommend WB for routine diagnosis of histoplasmosis by qPCR in patients with progressive disseminated histoplasmosis.
Título traducido de la contribuciónAplicación de ensayos de PCR en tiempo real para el diagnóstico de la histoplasmosis utilizando tres dianas moleculares en tejidos humanos de FFPE y sangre.
IdiomaEnglish (US)
PáginasS63
Número de páginas1
EstadoPublished - jun 2018
EventoXI Encuentro Nacional de Investigadores en Enfermedades Infecciosas - Pereira
Duración: ago 2 2018ago 4 2018

Conference

ConferenceXI Encuentro Nacional de Investigadores en Enfermedades Infecciosas
PaísColombia
CiudadPereira
Período8/2/188/4/18

Citar esto

López, L. F., Clay, O. K., Cáceres, D. H., & Gómez, B. L. (2018). Application of real-time PCR assays for the diagnosis of histoplasmosis using three molecular targets in human FFPE tissues and whole-blood. S63. Resumen desde XI Encuentro Nacional de Investigadores en Enfermedades Infecciosas, Pereira, .
López, Luisa F. ; Clay, Oliver Keatinge ; Cáceres, Diego H. ; Gómez, Beatriz L. / Application of real-time PCR assays for the diagnosis of histoplasmosis using three molecular targets in human FFPE tissues and whole-blood. Resumen desde XI Encuentro Nacional de Investigadores en Enfermedades Infecciosas, Pereira, .1 p.
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title = "Application of real-time PCR assays for the diagnosis of histoplasmosis using three molecular targets in human FFPE tissues and whole-blood",
abstract = "Objective: Histoplasmosis is a fungal infection that causes significantmorbidity andmortality in persons living withHIV/AIDS, especially in countries with limited resources. Currently used diagnostic tests rely on culture and serology, lack sensitivity and often require weeks to obtain results causing significant diagnosis delays; molecular assays are not commercially available. Methods: we aimed to apply quantitative real-time PCR (qPCR) targeting three protein-coding genes of Histoplasma capsulatum (100-kDa, H and M antigens) for detection of H. capsulatum infection in formalin-fixed paraffin-embedded (FFPE) and whole blood (WB) samples from patients with proven histoplasmosis. Results: For FFPE samples, the sensitivity of 100-kDa, H and M qPCR assays were 93.9{\%}, 91{\%} and 57{\%}, respectively; however, the same qPCR assays showed only 23{\%}, 19{\%} and 11.5{\%} of sensitivity for 100-kDa, H y M qPCR assays when used with the WB samples. The specificity of qPCR was determined by testing samples from patients with other clinical infections and healthy controls and was 93{\%}-100{\%} depending upon the assay and the specimen type. Conclusion: we applied three qPCR assays for detecting H. capsulatum DNA in human samples, and demonstrated that the molecular protocols based on amplification of 100-kDa and H antigen can be successfully used for diagnosing this mycosis when using FFPE samples; however, we do not recommend WB for routine diagnosis of histoplasmosis by qPCR in patients with progressive disseminated histoplasmosis.",
author = "L{\'o}pez, {Luisa F.} and Clay, {Oliver Keatinge} and C{\'a}ceres, {Diego H.} and G{\'o}mez, {Beatriz L.}",
year = "2018",
month = "6",
language = "English (US)",
pages = "S63",
note = "XI Encuentro Nacional de Investigadores en Enfermedades Infecciosas ; Conference date: 02-08-2018 Through 04-08-2018",

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Application of real-time PCR assays for the diagnosis of histoplasmosis using three molecular targets in human FFPE tissues and whole-blood. / López, Luisa F.; Clay, Oliver Keatinge; Cáceres, Diego H.; Gómez, Beatriz L.

2018. S63 Resumen desde XI Encuentro Nacional de Investigadores en Enfermedades Infecciosas, Pereira, .

Resultado de la investigación: Tipos de Contribuciónes en ConferenciaResumen

TY - CONF

T1 - Application of real-time PCR assays for the diagnosis of histoplasmosis using three molecular targets in human FFPE tissues and whole-blood

AU - López, Luisa F.

AU - Clay, Oliver Keatinge

AU - Cáceres, Diego H.

AU - Gómez, Beatriz L.

PY - 2018/6

Y1 - 2018/6

N2 - Objective: Histoplasmosis is a fungal infection that causes significantmorbidity andmortality in persons living withHIV/AIDS, especially in countries with limited resources. Currently used diagnostic tests rely on culture and serology, lack sensitivity and often require weeks to obtain results causing significant diagnosis delays; molecular assays are not commercially available. Methods: we aimed to apply quantitative real-time PCR (qPCR) targeting three protein-coding genes of Histoplasma capsulatum (100-kDa, H and M antigens) for detection of H. capsulatum infection in formalin-fixed paraffin-embedded (FFPE) and whole blood (WB) samples from patients with proven histoplasmosis. Results: For FFPE samples, the sensitivity of 100-kDa, H and M qPCR assays were 93.9%, 91% and 57%, respectively; however, the same qPCR assays showed only 23%, 19% and 11.5% of sensitivity for 100-kDa, H y M qPCR assays when used with the WB samples. The specificity of qPCR was determined by testing samples from patients with other clinical infections and healthy controls and was 93%-100% depending upon the assay and the specimen type. Conclusion: we applied three qPCR assays for detecting H. capsulatum DNA in human samples, and demonstrated that the molecular protocols based on amplification of 100-kDa and H antigen can be successfully used for diagnosing this mycosis when using FFPE samples; however, we do not recommend WB for routine diagnosis of histoplasmosis by qPCR in patients with progressive disseminated histoplasmosis.

AB - Objective: Histoplasmosis is a fungal infection that causes significantmorbidity andmortality in persons living withHIV/AIDS, especially in countries with limited resources. Currently used diagnostic tests rely on culture and serology, lack sensitivity and often require weeks to obtain results causing significant diagnosis delays; molecular assays are not commercially available. Methods: we aimed to apply quantitative real-time PCR (qPCR) targeting three protein-coding genes of Histoplasma capsulatum (100-kDa, H and M antigens) for detection of H. capsulatum infection in formalin-fixed paraffin-embedded (FFPE) and whole blood (WB) samples from patients with proven histoplasmosis. Results: For FFPE samples, the sensitivity of 100-kDa, H and M qPCR assays were 93.9%, 91% and 57%, respectively; however, the same qPCR assays showed only 23%, 19% and 11.5% of sensitivity for 100-kDa, H y M qPCR assays when used with the WB samples. The specificity of qPCR was determined by testing samples from patients with other clinical infections and healthy controls and was 93%-100% depending upon the assay and the specimen type. Conclusion: we applied three qPCR assays for detecting H. capsulatum DNA in human samples, and demonstrated that the molecular protocols based on amplification of 100-kDa and H antigen can be successfully used for diagnosing this mycosis when using FFPE samples; however, we do not recommend WB for routine diagnosis of histoplasmosis by qPCR in patients with progressive disseminated histoplasmosis.

M3 - Abstract

SP - S63

ER -

López LF, Clay OK, Cáceres DH, Gómez BL. Application of real-time PCR assays for the diagnosis of histoplasmosis using three molecular targets in human FFPE tissues and whole-blood. 2018. Resumen desde XI Encuentro Nacional de Investigadores en Enfermedades Infecciosas, Pereira, .