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

Exp Parasitol. 2020 Nov:218:107981. doi: 10.1016/j.exppara.2020.107981. Epub 2020 Aug 29.

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.

Keywords: Commercial extraction kits; In house extraction protocols; Molecular analysis; PCR; Sand fly; Vector.

MeSH terms

  • Analysis of Variance
  • Animals
  • Costs and Cost Analysis
  • DNA / analysis
  • DNA / isolation & purification*
  • DNA / standards
  • Electrophoresis, Agar Gel
  • Female
  • Male
  • Polymerase Chain Reaction / methods
  • Polymerase Chain Reaction / standards
  • Psychodidae / genetics*
  • Sodium Chloride
  • Spectrophotometry
  • Time Factors

Substances

  • Sodium Chloride
  • DNA