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

Author(s)

Franco Leonel, Joao Augusto ^{[1, 2]} ; Vioti, Geovanna ^{[1, 2]} ; Alves, Maria Luana ^{[1, 2]} ; da Silva, Diogo Tiago ^{[1, 2]} ; Meneghesso, Pedro Armando ^[1] ; Benassi, Julia Cristina ^[1] ; Pereira Spada, Julio Cesar ^{[1, 2]} ; Galvis-Ovallos, Fredy ^[3] ; Soares, Rodrigo Martins ^{[2, 4]} ; Ferreira de Sousa Oliveira, Tricia Maria ^{[1, 2]}

Affiliation

^[1] Univ Sao Paulo, Fac Anim Sci & Food Engn, Dept Vet Med, Lab Appl Prevent Vet Med, Pirassununga, SP - Brazil

^[2] Univ Sao Paulo, Fac Vet Med & Anim Sci, Dept Prevent Vet Med & Anim Hlth, Postgrad Program Expt Epidemiol Appl Zoonoses, Pirassununga, SP - Brazil

^[3] Univ Sao Paulo, Sch Publ Hlth, Lab Entomol Publ Hlth, Sao Paulo, SP - Brazil

^[4] Univ Sao Paulo, Fac Vet Med & Anim Sci, Dept Prevent Vet Med & Anim Hlth, Pirassununga, SP - Brazil

• Document type: Journal article
• Source: Experimental Parasitology; v. 218, NOV 2020.
• Web of Science Citations: 0

Abstract

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. (AU)

• FAPESP's process: 17/23436-0 - Hosts of Lutzomyia longipalpis in endemic area for visceral leishmaniasis
• Grantee: João Augusto Franco Leonel
• Support Opportunities: Scholarships in Brazil - Master