PCR-based detection of Leishmania DNA in skin samples of post kala-azar dermal leishmaniasis patients from an endemic area of Bangladesh.
Post kala-azar dermal leishmaniasis (PKDL) is a sequel of visceral leishmaniasis (VL) and PKDL patients  are an important reservoir for anthroponotic transmission of VL. Therefore, diagnosis and treatment of PKDL is  important for the kala-azar elimination program in South Asia, including Bangladesh. While definitive diagnosis  of PKDL is still based on microscopy, despite the low sensitivity of this method of diagnosis, PCR for  identification of kinetoplast DNA (kDNA) from Leishmania parasites is expected to be a rapid and  sensitive diagnostic method. We attempted PCR-based diagnosis from skin biopsy specimens and compared PCR to  other available detection methods in order to determine the acceptability and feasibility of the PCR diagnostic  method in an endemic area of VL in Bangladesh. Both skin biopsy specimens and blood samples were collected from  110 patients suspected to have PKDL from 6 subdistrict health complexes in Mymensingh, Bangladesh. Using  microscopy, we identified 32 samples (29.1%) that were positive for Leishmania. Immunochromatography  tests indicated that 85 samples (77.3%) were positive for Leishmania. In contrast, a total of 104 (94.5%)  samples tested positive using nested PCR, while unaffected portions of skin from PKDL patients tested negative.  Sequencing analysis of the PCR products indicated that the amplified portion had more than 98% nucleotide  sequence identity to the Leishmania donovani reference strain, D10. These findings indicate that the PCR  method using a skin biopsy is highly sensitive and useful for confirmatory diagnosis of PKDL.
Department of Microbiology, Mymensingh Medical College, Mymensingh, Bangladesh.
Syeda Anjuman
SA
Nasreen
Md Akram
MA
Hossain
Shyamal Kumar
SK
Paul
Md Chand
MC
Mahmud
Salma
S
Ahmed
Souvik
S
Ghosh
Nobumichi
N
Kobayashi
D001459:Bangladesh / Q000453:epidemiology; D004269:DNA, Bacterial; D058872:Epidemics; D006801:Humans; D007893:Leishmania donovani / Q000235:genetics* / Q000302:isolation & purification; D016773:Leishmaniasis, Cutaneous / Q000150:complications / Q000175:diagnosis* / Q000453:epidemiology; D007898:Leishmaniasis, Visceral / Q000150:complications; D016133:Polymerase Chain Reaction
D004269:DNA, Bacterial