Application of automated magnetic beads technology for DNA extraction in different vector arthropods

Molecular Biology has proven to be an essential tool in public health laboratories, as it provides significant advances in diagnosing and treating diseases and plays a crucial role in preventing and controlling epidemics. Considering the need to detect etiological agents from various vectors in the health service to improve knowledge of the regional transmission cycles of diseases involving arthropods, molecular analysis is an alternative for rapidly diagnosing natural infection by these organisms. This study aims to standardize commercial automated DNA extraction kits to obtain the DNA of pathogens from different vectors. Specimens of sandflies, ticks, fleas and kissing bugs received at LACEN-RO were screened and used in the tests. Three kits were evaluated: (i) Extracta Kit DNA and RNA from Pathogens (MPTA-PU16-B); (ii) Extracta Kit DNA from Bacteria (MBXD-PU16-B); and (iii) Extracta Kit DNA and RNA from Pathogens GOLD (MVXA-PU96 B/W Gold C). The specific protocol was used for each kit, and the EXTRACTA® 32 device was used to perform the automated extraction, following the manufacturer's instructions. PCRs were performed with universal primers for the mitochondrial DNA cytochrome c oxidase subunit I (COI) region and specific primers for Rickettsia spp., Leishmania spp. and Trypanosoma cruzi. The PCR products were detected using agarose gel electrophoresis, which enabled observing the amplified DNA, demonstrating the success of automated vector DNA extraction technique. These findings will enable future epidemiological studies to more quickly and effectively identify vectors and their parasites involved in the epidemiological cycles of diseases which occur in Rondônia state (Northern Brazil).