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Genetic fingerprinting and identification of differentially expressed genes in isolates of Leishmania donovani from Indian patients of post-kala-azar dermal leishmaniasis

Published online by Cambridge University Press:  28 August 2007

B. V. SUBBA RAJU ,
R. SINGH ,
G. SREENIVAS ,
S. SINGH  and
P. SALOTRA
B. V. SUBBA RAJU
Affiliation:
Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi-110 029, India
R. SINGH
Affiliation:
Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi-110 029, India
G. SREENIVAS
Affiliation:
Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi-110 029, India
S. SINGH
Affiliation:
Department of Microbiology, Jiwaji University, Gwalior, M.P.India
P. SALOTRA*
Affiliation:
Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi-110 029, India
*
*Corresponding author: Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi-110 029, India. Tel: +91 11 26166124. Fax: +91 11 26166124. E-mail: salotra@vsnl.com
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Summary

Post-kala-azar dermal leishmaniasis (PKDL) is an unusual dermatosis that develops as a sequel in 5–15% of cured cases of kala-azar (KA) after months or years of treatment in India. Molecular differences are reported to exist between the KA and PKDL isolates which may underlie the diversity in clinical manifestations of the disease. Here, arbitrary primed-PCR (AP-PCR) has been used for genetic fingerprinting of parasite isolates from dermal lesions of PKDL patients (n=14) and compared with bone-marrow derived parasites from KA patients (n=3). All isolates showed an identical AP-PCR pattern with 4 arbitrary primers. Further, AP-PCR was exploited to identify the stage regulated genes of the parasite. Six polymorphic fragments were identified in PKDL in comparison with KA isolates, and were subjected to Northern blot analysis. Five polymorphic fragments represented transcribed sequences; 4 out of 5 drew differential expression in pro- and amastigote stages, although the expression was comparable between PKDL and KA isolates. The study led to the identification of genes, which exhibit stage-regulated expression in Leishmania donovani derived from PKDL or KA patients, including a putative phosphodiesterase, DEAD box RNA helicase, iron superoxide dismutase b (fesodb) and a hypothetical protein. Demonstration of transcripts of DEAD box RNA helicase in PKDL and KA diseased tissues implicates its role in disease pathogenesis.

Keywords

Leishmania donovaniPKDLvisceral leishmaniasisAPPCRdifferentially expressed genesgenomic fingerprinting
Type
Research Article
Information
Parasitology , Volume 135 , Issue 1 , January 2008 , pp. 23 - 32
Copyright
Copyright © Cambridge University Press 2007

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