DNA extraction from individual Phlebotomine sand flies (Diptera: Psychodidae: Phlebotominae) specimens: Which is the method with better results?

Phlebotomine sand flies (Diptera: Psychodidae: Phlebotominae) are a group of small insects of great concern for Public Health. These dipterous are intensely studied worldwide due to their involvement in the transmission of several pathogens, mainly Leishmania spp. parasites. Nowadays, the molecular...

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Published inExperimental parasitology Vol. 218; p. 107981
Main Authors Leonel, João Augusto Franco, Vioti, Geovanna, Alves, Maria Luana, da Silva, Diogo Tiago, Meneghesso, Pedro Armando, Benassi, Julia Cristina, Spada, Julio Cesar Pereira, Galvis-Ovallos, Fredy, Soares, Rodrigo Martins, Oliveira, Trícia Maria Ferreira de Sousa
Format Journal Article
LanguageEnglish
Published Elsevier Inc 01.11.2020
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Summary:Phlebotomine sand flies (Diptera: Psychodidae: Phlebotominae) are a group of small insects of great concern for Public Health. These dipterous are intensely studied worldwide due to their involvement in the transmission of several pathogens, mainly Leishmania spp. parasites. Nowadays, the molecular tools have been included in Phlebotomine sand flies studies and has shown to be powerful tools in bioecology studies of these dipterous. Thereby, when molecular approaches are employed, there is a great concern regarding the amount and quality of the DNA obtained for analysis. Here, seven methods of DNA extraction, between commercial kits and in house extraction protocols were evaluated. We considered measure of DNA concentration and purity ratios using a spectrophotometer to check the performance of each protocol. In addition, the quality evaluation of the DNA extracted was performed by endogenous gene PCR on samples. The results of the seven evaluated DNA extraction protocols and their implications are discussed. [Display omitted] •In house or commercial DNA extraction methods were able to extract DNA from individual Phlebotomine sand flies samples.•In house extraction methods performed better or at least equal to the commercial methods.•The in house method based on NaCl/alcohol is a good and safe choice to extract DNA from Phlebotomine sand flies.
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ISSN:0014-4894
1090-2449
DOI:10.1016/j.exppara.2020.107981