The identification of sandfly species, from an area of Argentina with endemic leishmaniasis, by the PCR-based analysis of the gene coding for 18S ribosomal RNA

Abstract

The area around Río Blanco, in the Orán department in the north of the Argentinian province of Salta, is endemic for American tegumentary leishmaniasis. In an attempt to facilitate the identification of the Lutzomyia species in this area, sequences of the gene coding for the 18S ribosomal RNA (rRNA) of sandflies caught in a Shannon trap were explored, by a combination of PCR and analysis of restriction-fragment-length polymorphism (RFLP). The products from the PCR, which employed two primers developed specifically for this study (Lu.18S 1S and Lu.18S AR), were cloned into a commercial vector (pGEM-T Easy) so that their nucleotide sequences could be investigated. In the RFLP analysis, the products of single and double digestion with the AfaI and HapII restriction enzymes were separated by electrophoresis in 3% or 4% agarose. Taken together with the results of a morphological investigation of the flies, the resultant DNA fragment patterns were sufficient to identify most of the sandflies caught as Lu. neivai. Although two other species, Lu. cortelezzii and Lu. sallesi, were collected, they were relatively rare and only identified morphologically. A single digestion of the 18S-rRNA gene sequences with AfaI or HapII appeared sufficient and useful for the identification of Lu. neivai from the north of Salta province, and for several other Lutzomyia species.

Endemic Transmission of Visceral Leishmaniasis in Bhutan

“…14 The 18SrRNA gene from female sand flies was amplified using the primer pair Lu.18S 1S (5 -TGCCAGTAGTTAT ATGCTTG-3 )/Lu.18S AR (5 -CACCTACGGAAACCTTG TTAC-3 ) as described elsewhere. 15 The PCR products were run on 1.5% agarose gels stained with ethidium bromide and visualized under UV light.…”

Phylogenic analysis of Chinese Leishmania isolates based on small subunit ribosomal RNA (SSU rRNA) and 7 spliced leader RNA (7SL RNA)

“…A simpler and more discriminatory method is required. Genetic markers have been explored for the development of reliable tools for species identification as well as an understanding of intraspecies genetic diversity and population structure (Aransay et al 1999;Barroso et al 2007;Barón et al 2008).…”

Urban Transmission of American Cutaneous Leishmaniasis in Argentina: Spatial Analysis Study

“…punctigeniculata . [5][6][7][8] Lutzomyia neivai is the unique species of phlebotomus sand fly in the region for which infection with Leishmania sp. has been proved.…”