Journal Article

Detection and Species Identification of <i>Leishmania</i> DNA from Filter Paper Lesion Impressions for Patients with American Cutaneous Leishmaniasis

Andrea K. Boggild, Braulio Mark Valencia, Diego Espinosa, Nicolas Veland, Ana Pilar Ramos, Jorge Arevalo, Alejandro Llanos-Cuentas and Donald E. Low

in Clinical Infectious Diseases

Published on behalf of Infectious Diseases Society of America

Volume 50, issue 1, pages e1-e6
Published in print January 2010 | ISSN: 1058-4838
Published online January 2010 | e-ISSN: 1537-6591 | DOI: http://dx.doi.org/10.1086/648730
Detection and Species Identification of Leishmania DNA from Filter Paper Lesion Impressions for Patients with American Cutaneous Leishmaniasis

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Background

Traditional detection of Leishmania from ulcers involves collection of invasive specimens that cause discomfort, require technical expertise, and carry risks of invasive procedures. We compared traditional diagnostic methods with a molecular noninvasive filter paper-based method for the diagnosis of cutaneous leishmaniasis.

Methods

Consecutive patients presenting to the Leishmania Clinic at Hospital Nacional Cayetano Heredia were enrolled. Polymerase chain reaction (PCR) was performed on lesion scrapings, aspirates, and filter paper impressions. The reference standard was any 2 of 5 tests positive: smear, aspirate culture, invasive-specimen PCR (scrapings and aspirates), filter paper PCR, and leishmanin skin test. Outcome measures were sensitivity and specificity. Leishmania speciation was performed by PCR-restriction fragment length polymorphism (RFLP) of positive specimens.

Results

Forty-five patients with 66 lesions were enrolled. Of 52 lesions diagnosed as cutaneous leishmaniasis, 50 were positive by PCR of invasive specimens versus 48 by PCR of filter papers (P = .930). Sensitivity and specificity of PCR on invasively obtained specimens were 94.2% (95% confidence interval [CI], 87.9%–100%) and 92.9% (95% CI, 79.4%–100%). Sensitivity and specificity of filter paper PCR were 92.3% (95% CI, 85.1%–99.5%) and 100%. Culture, smear, and leishmanin skin test all had inferior sensitivities, compared with PCR of invasive or noninvasive specimens (P < .001). Of 50 specimens positive by PCR, 19 had sufficient DNA for PCR-RFLP analysis.

Conclusions

Filter paper PCR constitutes a sensitive and specific alternative to traditional diagnostic assays. This novel, rapid, well-tolerated method has the potential for widespread use in the field and in pediatric populations where traditional specimen collection is most difficult to perform, and can potentially be used for rapid species identification.

Journal Article.  4151 words.  Illustrated.

Subjects: Infectious Diseases ; Immunology ; Public Health and Epidemiology ; Microbiology

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