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

Título traducido de la contribución: 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.
Idioma originalEnglish (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, .