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A real-time PCR assay to estimate Leishmania chagasi load in its natural sand fly vector Lutzomyia longipalpis

Ranasinghe, S., Rogers, Matthew E., Hamilton, J. G. C., Bates, Paul and Maingon, R. D. C. (2008) 'A real-time PCR assay to estimate Leishmania chagasi load in its natural sand fly vector Lutzomyia longipalpis'. Transactions of the Royal Society of Tropical Medicine and Hygiene, Vol 102, Issue 9, pp. 875-882.

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Abstract

Leishmania chagasi, transmitted mainly by Lutzomyia longipalpis sand flies, causes visceral leishmaniasis and atypical cutaneous leishmaniasis in Latin America. Successful vector control depends upon determining vectorial capacity and understanding Leishmania transmission by sand flies. As microscopic detection of Leishmania in dissected sand fly guts is laborious and time-consuming, highly specific, sensitive, rapid and robust Leishmania PCR assays have attracted epidemiologists' attention. Real-time PCR is faster than qualitative PCR and yields quantitative data amenable to statistical analyses. A highly reproducible Leishmania DNA polymerase gene-based TaqMan real-time PCR assay was adapted to quantify Leishmania in sand flies, showing intra-assay and inter-assay coefficient variations lower than 1 and 1.7%, respectively, and sensitivity to 10 pg Leishmania DNA (similar to 120 parasites) in as much as 100 ng sand fly DNA. Data obtained for experimentally infected sand flies yielded parasite loads within the range of counts obtained by microscopy for the same sand fly cohort or that were around five times higher than microscopy counts, depending on the method used for data analysis. These results highlight the potential of quantitative PCR for Leishmania transmission studies, and the need to understand factors affecting its sensitivity and specificity. (C) 2008 Royal Society of Tropical Medicine and Hygiene. Published by Elsevier Ltd. All rights reserved.

Item Type: Article
Uncontrolled Keywords: leishmania sand fly pcr sensitivity and specificity transmission epidemiology polymerase-chain-reaction visceral leishmaniasis phlebotomus-papatasi infantum DNA transmission flies gene quantification identification infection
Subjects: QX Parasitology > Insects. Other Parasites > QX 505 Diptera
WC Communicable Diseases > Tropical and Parasitic Diseases > WC 755 Epidemiology
WR Dermatology > Parasitic Skin Diseases > WR 350 Tropical diseases of the skin. Mucocutaneous leishmaniasis. Leishmaniasis
WC Communicable Diseases > Tropical and Parasitic Diseases > WC 715 Visceral leishmaniasis
QU Biochemistry > Genetics > QU 470 Genetic structures
Faculty: Department: Groups (2002 - 2012) > Molecular & Biochemical Parasitology Group
Digital Object Identifer (DOI): https://doi.org/10.1016/j.trstmh.2008.04.003
Depositing User: Mary Creegan
Date Deposited: 19 Jul 2010 13:23
Last Modified: 06 Feb 2018 13:00
URI: http://archive.lstmed.ac.uk/id/eprint/901

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