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Sensitive diagnosis of cutaneous leishmaniasis by lesion swab sampling coupled to qPCR

Published online by Cambridge University Press:  11 August 2014

EMILY R. ADAMS ,
MARIA ADELAIDA GOMEZ ,
LAURA SCHESKE ,
RUBY RIOS ,
RICARDO MARQUEZ ,
ALEXANDRA COSSIO ,
AUDREY ALBERTINI ,
HENK SCHALLIG  and
NANCY GORE SARAVIA
EMILY R. ADAMS*
Affiliation:
Department of Parasitology, Royal Tropical Institute, Meibergdreef 39, 1105 AZ Amsterdam, The Netherlands Department of Parasitology, Liverpool School of Tropical Medicine, Pembroke Place, Liverpool L3 5QA, UK
MARIA ADELAIDA GOMEZ
Affiliation:
Centro Internacional de Entrenamiento e Investigaciones Médicas (CIDEIM), Cra 125 # 19-225, Cali, Colombia
LAURA SCHESKE
Affiliation:
Department of Parasitology, Royal Tropical Institute, Meibergdreef 39, 1105 AZ Amsterdam, The Netherlands
RUBY RIOS
Affiliation:
Centro Internacional de Entrenamiento e Investigaciones Médicas (CIDEIM), Cra 125 # 19-225, Cali, Colombia
RICARDO MARQUEZ
Affiliation:
Centro Internacional de Entrenamiento e Investigaciones Médicas (CIDEIM), Cra 125 # 19-225, Cali, Colombia
ALEXANDRA COSSIO
Affiliation:
Centro Internacional de Entrenamiento e Investigaciones Médicas (CIDEIM), Cra 125 # 19-225, Cali, Colombia
AUDREY ALBERTINI
Affiliation:
Foundation for Innovative New Diagnostics, Geneva, Switzerland
HENK SCHALLIG
Affiliation:
Department of Parasitology, Royal Tropical Institute, Meibergdreef 39, 1105 AZ Amsterdam, The Netherlands
NANCY GORE SARAVIA
Affiliation:
Centro Internacional de Entrenamiento e Investigaciones Médicas (CIDEIM), Cra 125 # 19-225, Cali, Colombia
*
*Corresponding author: Department of Parasitology, Liverpool School of Tropical Medicine, Pembroke Place, Liverpool L3 5QA, UK. E-mail: e.adams@liv.ac.uk
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Summary

Variation in clinical accuracy of molecular diagnostic methods for cutaneous leishmaniasis (CL) is commonly observed depending on the sample source, the method of DNA recovery and the molecular test. Few attempts have been made to compare these variables. Two swab and aspirate samples from lesions of patients with suspected CL (n = 105) were evaluated alongside standard diagnosis by microscopic detection of amastigotes or culture of parasites from lesion material. Three DNA extraction methods were compared: Qiagen on swab and aspirate specimens, Isohelix on swabs and Boil/Spin of lesion aspirates. Recovery of Leishmania DNA was evaluated for each sample type by real-time polymerase chain reaction detection of parasitic 18S rDNA, and the diagnostic accuracy of the molecular method determined. Swab sampling combined with Qiagen DNA extraction was the most efficient recovery method for Leishmania DNA, and was the most sensitive (98%; 95% CI: 91–100%) and specific (84%; 95% CI: 64–95%) approach. Aspirated material was less sensitive at 80% (95% CI: 70–88%) and 61% (95% CI: 50–72%) when coupled to Qiagen or Boil-Spin DNA extraction, respectively. Swab sampling of lesions was painless, simple to perform and coupled with standardized DNA extraction enhances the feasibility of molecular diagnosis of CL.

Keywords

swab samplingqPCRcutaneous leishmaniasisdiagnosismolecular diagnosis
Type
Special Issue Article
Information
Parasitology , Volume 141 , Issue 14: Symposia of the British Society for Parasitology Volume 52 Advances in diagnostics for parasitic diseases , December 2014 , pp. 1891 - 1897
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Copyright
Copyright © Cambridge University Press 2014 

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