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Molecular Characterization Of Leishmania Infection In Sand Flies From Al-madinah Province, Western Saudi Arabia | 2791
Journal of Community Medicine & Health Education
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Cutaneous leishmaniasis (CL) is caused by various species of the genus Leishmania. The disease is considered a major health
problem in different areas of Saudi Arabia including Al-madinah Al-munawarah province. We aimed to identify Leishmania
species isolated from sand fly vectors by molecular analysis. Sand fly sampling was carried out from May 2010-October 2010
in province of Al-madinah Al-munawarah from four different localities. Female sand flies collected were subjected to DNA
extraction followed by molecular analysis using the nested PCR and conventional PCR protocols, respectively, against minicircle
kDNA and ribosomal internal transcribed spacer 1 (ITS1-rDNA). The PCR positive specimens against ITS1-rDNA locus were
digested for further confirmation of species identification. A total of 2910 sand flies were collected. P. papatasi accounted for
93.8% (1673 male and 1057 female), however, the number of P. sergenti was only 180 (109 male and 71 female). Sixty-two out
of 250 (23.7%) female P. papatasi tested for Leishmania parasite were positive for L. major using the semi-nested PCR method
against kDNA. All of the 62 positive specimens produced a band size 650bp. A 31% of female P. sergenti were positive against
kDNA of L. tropica and produced a 720bp band. These positive P.sergenti for L. tropica DNA produced ITS1-PCR-RFLP profile
showed two bands of~200bp and 57bp which are specific for L. tropica, confirming the presence of L. tropica in P. sergenti.
However, the ITS1-PCR-RFLP profile showed two bands of~203bp and 132bp which are specific for L. major in P. papatasi. We
concluded that, the semi-nested PCR method against kDNA and the ITS1-PCR-RFLP analysis are useful tools for molecular
identification of both L. major and L. tropica. A multicenter study is necessary in order to evaluate the extent of the disease and
functional analysis of new Leishmania genes.
Acknowledgements: This study was supported by grant funded by the Strategic Science, Advanced Technologies and Innovation
Unit (SRU)-Taibah University-Al-madinah Al-munawarah, Saudi Arabia (grant number 08-BIO25-5).
Professor and Consultant of Medical Biochemistry and Molecular Biology and Head of Medical Laboratories Technology Department, Faculty of
Applied Medical Sciences Taibah University, Saudi Arabia.
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