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Journal of Clinical Microbiology, February 2002, p. 601-606, Vol. 40, No. 2
0095-1137/01/$04.00+0     DOI: 10.1128/JCM.40.2.601-606.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

PCR-Based Diagnosis of Acute and Chronic Cutaneous Leishmaniasis Caused by Leishmania (Viannia)

Kristen A. Weigle,1 Luz Angela Labrada,2* Caterin Lozano,2 Cecilia Santrich,2 and Douglas C. Barker3

Department of Epidemiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina,1 Corporacion CIDEIM, Cali, Colombia,2 MRC Outstation of NIMR, Molteno Laboratories, University of Cambridge, Cambridge, United Kingdom3

Received 4 June 2001/ Returned for modification 23 September 2001/ Accepted 2 December 2001

We evaluated PCR methods for diagnosis of acute and chronic cutaneous leishmaniasis (CL) in an area of Colombia where Leishmania (Viannia) is endemic. The PCR method specifically amplified whole linearized minicircle kinetoplast DNA (kDNA) of the Leishmania subgenus Viannia from biopsy lysates. PCR products were detected in agarose gels. For 255 acute cases, this PCR method had greater sensitivity (75.7%) than each conventional method, i.e., microscopic examination of Giemsa-stained lesion scraping (46.7%), biopsy culture (55.3%), aspirate culture (46.3%), and the conventional methods combined (70.2%). Among 44 cases of chronic CL, amplification of biopsy DNA was more sensitive (45.5%) than the individual (4.5 to 27.7%) and combined (27.3%) conventional methods. The detection of kDNA in biopsies from chronic lesions was enhanced by a chemiluminescent dot blot hybridization, which produced a sensitivity of 65.8% when alone and 90.9% when in combination with DNA extraction of biopsy lysates (P < 0.001). Three biopsies from 84 skin lesions of other etiologies were falsely positive by PCR (specificity, 96.4%). PCR detected kDNA more frequently in biopsies (detection level, 83.9%) than in aspirates (74.7%) from 103 cases of acute CL. Among aspirates from 53 chronic cases of CL, the alternative methods, DNA extraction and hybridization, increased sensitivity from 41.5 to 56.6% (P > 0.05). This enhanced PCR method in chronic biopsies was so much more sensitive than conventional methods that it should be considered the preferred diagnostic method for chronic CL. These findings support the appropriate incorporation of PCR into diagnostic strategies for cutaneous leishmaniasis.

* Corresponding author. Mailing address: Corporacion CIDEIM, Avenida 1N #3-03, Centenario, Apartado Aéreo 5390, Cali, Colombia. Phone: 57 2 5581931. Fax: 57 2 6672989. E-mail: luzlabrada{at}hotmail.com.

Journal of Clinical Microbiology, February 2002, p. 601-606, Vol. 40, No. 2
0095-1137/01/$04.00+0     DOI: 10.1128/JCM.40.2.601-606.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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