A real-time PCR assay for detection and quantification of Leishmania (Viannia) in skin and mucosal lesions: an exploratory study of parasite load and clinical parameters

M. Jara; V. Adaui; B.M. Valencia; D. Martinez Medina; M. Alba; C. Castrillon; M. Cruz; I. Cruz; G. Van der Auwera; A. Llanos-Cuentas; J.C. Dujardin; J. Arevalo

doi:10.1128/JCM.00208-13

A real-time PCR assay for detection and quantification of Leishmania (Viannia) in skin and mucosal lesions: an exploratory study of parasite load and clinical parameters

M. Jara, V. Adaui, B.M. Valencia, D. Martinez Medina, M. Alba, C. Castrillon, M. Cruz, I. Cruz, G. Van der Auwera, A. Llanos-Cuentas, J.C. Dujardin, J. Arevalo

Molecular Parasitology

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Abstract

Earlier histopathology studies suggest that parasite load may differ between cutaneous leishmaniasis (CL) and mucosal leishmaniasis (ML) lesions, and between acute and chronic CL. Formal demonstration requires highly sensitive detection and accurate quantification of Leishmania in human lesional tissue. In this study, we developed a quantitative real-time PCR (qPCR) assay targeting the minicircle kinetoplast DNA (kDNA) to detect and quantify Leishmania (Viannia) parasites. We evaluated a total of 156 lesion biopsies from CL or ML suspected cases, and compared the quantitative performance of our kDNA-qPCR assay with a previously validated qPCR assay based on the glucose-6-phosphate dehydrogenase (G6PD) gene. We also examined the relationship between parasite load and clinical parameters. The kDNA-qPCR sensitivity for Leishmania detection was 97.9% and its specificity was 87.5%. The parasite loads quantified by kDNA-qPCR and G6PD-qPCR assays were highly correlated (r=0.87, P

Original language	English
Journal	Journal of Clinical Microbiology
Volume	51
Issue number	6
Pages (from-to)	1826-1833
Number of pages	8
ISSN	0095-1137
DOIs	https://doi.org/10.1128/JCM.00208-13
Publication status	Published - 2013

Keywords

Protozoal diseases
Kala azar
Visceral
Cutaneous
Leishmaniasis
Leishmania donovani
Vectors
Sandflies
Phlebotomus argentipes
Detection
Quantification
Skin lesions
Mucosal
Parasite density
Clinical diagnosis
Real-time
Polymerase chain reaction
PCR
Assays
Biopsy
Kinetoplast DNA
Specificity
Sensitivity
Laboratory techniques and procedures

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10.1128/JCM.00208-13

2013jcmi1826

http://lib.itg.be/pdf/itg/2013/2013jcmi1826.pdf

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Jara, M., Adaui, V., Valencia, B. M., Martinez Medina, D., Alba, M., Castrillon, C., Cruz, M., Cruz, I., Van der Auwera, G., Llanos-Cuentas, A., Dujardin, J. C., & Arevalo, J. (2013). A real-time PCR assay for detection and quantification of Leishmania (Viannia) in skin and mucosal lesions: an exploratory study of parasite load and clinical parameters. Journal of Clinical Microbiology, 51(6), 1826-1833. https://doi.org/10.1128/JCM.00208-13

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title = "A real-time PCR assay for detection and quantification of Leishmania (Viannia) in skin and mucosal lesions: an exploratory study of parasite load and clinical parameters",

abstract = "Earlier histopathology studies suggest that parasite load may differ between cutaneous leishmaniasis (CL) and mucosal leishmaniasis (ML) lesions, and between acute and chronic CL. Formal demonstration requires highly sensitive detection and accurate quantification of Leishmania in human lesional tissue. In this study, we developed a quantitative real-time PCR (qPCR) assay targeting the minicircle kinetoplast DNA (kDNA) to detect and quantify Leishmania (Viannia) parasites. We evaluated a total of 156 lesion biopsies from CL or ML suspected cases, and compared the quantitative performance of our kDNA-qPCR assay with a previously validated qPCR assay based on the glucose-6-phosphate dehydrogenase (G6PD) gene. We also examined the relationship between parasite load and clinical parameters. The kDNA-qPCR sensitivity for Leishmania detection was 97.9% and its specificity was 87.5%. The parasite loads quantified by kDNA-qPCR and G6PD-qPCR assays were highly correlated (r=0.87, P",

keywords = "Protozoal diseases, Kala azar, Visceral, Cutaneous, Leishmaniasis, Leishmania donovani, Vectors, Sandflies, Phlebotomus argentipes, Detection, Quantification, Skin lesions, Mucosal, Parasite density, Clinical diagnosis, Real-time, Polymerase chain reaction, PCR, Assays, Biopsy, Kinetoplast DNA, Specificity, Sensitivity, Laboratory techniques and procedures",

author = "M. Jara and V. Adaui and B.M. Valencia and {Martinez Medina}, D. and M. Alba and C. Castrillon and M. Cruz and I. Cruz and {Van der Auwera}, G. and A. Llanos-Cuentas and J.C. Dujardin and J. Arevalo",

note = "FTX; ITG-B9A; ITG-B11A; DBM; U-MOLPAR; JIF; DOI; PDF; Abstract; DSPACE56",

year = "2013",

doi = "10.1128/JCM.00208-13",

language = "English",

volume = "51",

pages = "1826--1833",

journal = "Journal of Clinical Microbiology",

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publisher = "American Society for Microbiology",

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Jara, M, Adaui, V, Valencia, BM, Martinez Medina, D, Alba, M, Castrillon, C, Cruz, M, Cruz, I, Van der Auwera, G, Llanos-Cuentas, A, Dujardin, JC & Arevalo, J 2013, 'A real-time PCR assay for detection and quantification of Leishmania (Viannia) in skin and mucosal lesions: an exploratory study of parasite load and clinical parameters', Journal of Clinical Microbiology, vol. 51, no. 6, pp. 1826-1833. https://doi.org/10.1128/JCM.00208-13

A real-time PCR assay for detection and quantification of Leishmania (Viannia) in skin and mucosal lesions: an exploratory study of parasite load and clinical parameters. / Jara, M.; Adaui, V.; Valencia, B.M. et al.
In: Journal of Clinical Microbiology, Vol. 51, No. 6, 2013, p. 1826-1833.

Research output: Contribution to journal › A1: Web of Science-article › peer-review

TY - JOUR

T1 - A real-time PCR assay for detection and quantification of Leishmania (Viannia) in skin and mucosal lesions: an exploratory study of parasite load and clinical parameters

AU - Jara, M.

AU - Adaui, V.

AU - Valencia, B.M.

AU - Martinez Medina, D.

AU - Alba, M.

AU - Castrillon, C.

AU - Cruz, M.

AU - Cruz, I.

AU - Van der Auwera, G.

AU - Llanos-Cuentas, A.

AU - Dujardin, J.C.

AU - Arevalo, J.

N1 - FTX; ITG-B9A; ITG-B11A; DBM; U-MOLPAR; JIF; DOI; PDF; Abstract; DSPACE56

PY - 2013

Y1 - 2013

N2 - Earlier histopathology studies suggest that parasite load may differ between cutaneous leishmaniasis (CL) and mucosal leishmaniasis (ML) lesions, and between acute and chronic CL. Formal demonstration requires highly sensitive detection and accurate quantification of Leishmania in human lesional tissue. In this study, we developed a quantitative real-time PCR (qPCR) assay targeting the minicircle kinetoplast DNA (kDNA) to detect and quantify Leishmania (Viannia) parasites. We evaluated a total of 156 lesion biopsies from CL or ML suspected cases, and compared the quantitative performance of our kDNA-qPCR assay with a previously validated qPCR assay based on the glucose-6-phosphate dehydrogenase (G6PD) gene. We also examined the relationship between parasite load and clinical parameters. The kDNA-qPCR sensitivity for Leishmania detection was 97.9% and its specificity was 87.5%. The parasite loads quantified by kDNA-qPCR and G6PD-qPCR assays were highly correlated (r=0.87, P

AB - Earlier histopathology studies suggest that parasite load may differ between cutaneous leishmaniasis (CL) and mucosal leishmaniasis (ML) lesions, and between acute and chronic CL. Formal demonstration requires highly sensitive detection and accurate quantification of Leishmania in human lesional tissue. In this study, we developed a quantitative real-time PCR (qPCR) assay targeting the minicircle kinetoplast DNA (kDNA) to detect and quantify Leishmania (Viannia) parasites. We evaluated a total of 156 lesion biopsies from CL or ML suspected cases, and compared the quantitative performance of our kDNA-qPCR assay with a previously validated qPCR assay based on the glucose-6-phosphate dehydrogenase (G6PD) gene. We also examined the relationship between parasite load and clinical parameters. The kDNA-qPCR sensitivity for Leishmania detection was 97.9% and its specificity was 87.5%. The parasite loads quantified by kDNA-qPCR and G6PD-qPCR assays were highly correlated (r=0.87, P

KW - Protozoal diseases

KW - Kala azar

KW - Visceral

KW - Cutaneous

KW - Leishmaniasis

KW - Leishmania donovani

KW - Vectors

KW - Sandflies

KW - Phlebotomus argentipes

KW - Detection

KW - Quantification

KW - Skin lesions

KW - Mucosal

KW - Parasite density

KW - Clinical diagnosis

KW - Real-time

KW - Polymerase chain reaction

KW - PCR

KW - Assays

KW - Biopsy

KW - Kinetoplast DNA

KW - Specificity

KW - Sensitivity

KW - Laboratory techniques and procedures

U2 - 10.1128/JCM.00208-13

DO - 10.1128/JCM.00208-13

M3 - A1: Web of Science-article

C2 - 23554201

SN - 0095-1137

VL - 51

SP - 1826

EP - 1833

JO - Journal of Clinical Microbiology

JF - Journal of Clinical Microbiology

IS - 6

ER -

A real-time PCR assay for detection and quantification of Leishmania (Viannia) in skin and mucosal lesions: an exploratory study of parasite load and clinical parameters

Abstract

Keywords

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