Development of a Species-Specific PCR Assay for Detection of Leishmania donovani in Clinical Samples from Patients with Kala-Azar and Post-Kala-Azar Dermal Leishmaniasis
Open Access
- 1 March 2001
- journal article
- research article
- Published by American Society for Microbiology in Journal of Clinical Microbiology
- Vol. 39 (3), 849-854
- https://doi.org/10.1128/jcm.39.3.849-854.2001
Abstract
We have developed a PCR assay that is capable of amplifying kinetoplast DNA (kDNA) of Leishmania donovani in a species-specific manner among Old World leishmanias. With Indian strains and isolates of L. donovani the assay was sensitive enough to detect kDNA in an amount equivalent to a single parasite or less. The extreme sensitivity of the assay was reflected in its ability to detect parasite DNA from small volumes of peripheral blood of patients with kala-azar (KA) and from skin lesions from patients with post-KA dermal leishmaniasis (PKDL). A total of 107 clinical leishmaniasis samples were analyzed. Of these 102 (95.3%) were positive by PCR. The test provided a diagnosis of KA with 96% sensitivity using patient whole-blood samples instead of bone marrow or spleen aspirates that are obtained by invasive procedures. The assay was also successful in the diagnosis of 45 of 48 PKDL cases (93.8%). Cross-reactions with pathogens prevalent in the area of endemicity, viz., Mycobacterium tuberculosis, Mycobacterium leprae , and Plasmodium spp., could be ruled out. Eighty-one control samples, including dermal scrapings from healthy portions of skin from patients with PKDL were all negative. Two of twenty controls from the area of endemicity were found positive by PCR assay; however, there was a good possibility that these two were asymptomatic carriers since they were serologically positive for KA. Thus, this PCR assay represents a tool for the diagnosis of KA and PKDL in Indian patients in a noninvasive manner, with simultaneous species identification of parasites in clinical samples.Keywords
This publication has 35 references indexed in Scilit:
- Diagnosis of visceral leishmaniasis by the polymerase chain reaction using blood, bone marrow and lymph node samples from patients from the SudanTropical Medicine & International Health, 1997
- Diagnosis of visceral leishmaniasis by the polymerase chain reaction using blood, bone marrow and lymph node samples from patients from the SudanTropical Medicine & International Health, 1997
- Post‐Kala‐Azar Dermal Leishmaniasis in an HTV‐PatientInternational Journal of Dermatology, 1996
- rK39: A Cloned Antigen of Leishmania chagasi that Predicts Active Visceral LeishmaniasisThe Journal of Infectious Diseases, 1996
- Development of a genus specific primer set for detection ofLeishmaniaparasites by polymerase chain reactionFEMS Microbiology Letters, 1996
- Detection of Leishmania in the blood of early kala-azar patients with the aid of the polymerase chain reactionTransactions of the Royal Society of Tropical Medicine and Hygiene, 1995
- Diagnosis Of Symptomatic Visceral Leishmaniasis By Use Of The Polymerase Chain Reaction On Patient BloodThe Journal of Infectious Diseases, 1995
- Post‐kala‐azar dermal leishmaniasis: A light and electron microscopic study of 18 casesJournal of Cutaneous Pathology, 1993
- Cloning and characterization of differentially expressed genes from in vitro-grown ‘amastigotes’ of Leishmania donovaniMolecular and Biochemical Parasitology, 1993
- Rapid and sensitive detection of Leishmania kinetoplast DNA from spleen and blood samples of kala-azar patientsParasitology, 1992