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Centro de Investigaciones Regionales Dr. Hideyo Noguchi, Universidad Autónoma de Yucatán

2008, Number 1

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Rev Biomed 2008; 19 (1)

Design and evaluation of ITS2 PCR-RFLP based methods for molecular identification of mosquitoes Anopheles spp. (Diptera: Culicidae) from the Pacific Coast of Colombia

Cienfuegos AV, Gómez GF, Córdoba LA, Luckhart S, Conn JE, Correa MM

Language: Spanish
References: 34
Page: 35-44
PDF size: 518.86 Kb.


ABSTRACT

Introduction. Difficulties in anopheline identification due to intra-individual variation and interspecies similarity have led to the development of molecular tools that allow species discrimination.
Objective. To use bioinformatics tools to develop a PCR-RFLP assay for molecular identification of species belonging to the Oswaldoi Group reported in Colombia and to evaluate the usefulness of a PCR-RFLP-AluI assay previously developed in our lab, in anophelines from different Colombian localities.
Material and Methods. In silico restriction patterns were generated by bioinformatics analysis of ITS2 sequences reported in GenBank, corresponding to species belonging to the Oswaldoi Group. To evaluate the PCR-RFLP-AluI assay we used mosquitoes collected in four localities of the Pacific Coast of Colombia: Nuquí, Pizarro, Buenaventura and Tumaco.
Results. Bioinformatic analysis of reported ITS2 sequences showed that the enzyme AluI produced in silico restriction patterns that distinguished four species belonging to the Oswaldoi Group; two additional species could be differentiated with a second digest. The evaluation of the PCR-RFLP-AluI confirmed results of morphological identifications and demonstrated identification problems that can arise due to intraspecific variation and interspecies similarity.
Conclusions. The expansion of the PCR-RFLP-AluI methodology and its application in different regions of the country will allow rapid and accurate discrimination of a large number of mosquitoes, some of them belonging to species that are difficult to discriminate morphologically. This strategy will be useful for complementary studies to assess malaria transmission dynamics in Colombia.


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