| Congresso Brasileiro de Microbiologia 2023 | Resumo: 626-1 | ||||
Resumo:Currently, there is need to obtain economical, fast and efficient techniques for obtaining DNA, as techniques for identifying and detecting pathogens by PCR are becoming increasingly popular. Although PCR is a highly sensitive technique, it can require expensive reagents, specific equipment and considerable time for sample processing. Faced with these challenges, researches have developed alternative methods to speed up the process of extracting nucleic acids from bacteria, such as the use of rapid nucleic acid extraction reagents. Thus, this work aimed to evaluate a DNA extraction method by breaking the cell wall, using Pi-Lise Nucleic Acid Extraction Reagent. Suspensions of gram-positive bacteria, including Staphylococcus aureus ATCC 29213 and ATCC 33591, were used. The bacterial cells were centrifuged and the cell pellet was subjected to washing and solubilization steps with the rapid nuclei acid extraction reagent. The mixture was incubated at 95 °C in a dry bath for 5 minutes and, after centrifugation, the samples were submitted to conventional PCR analysis using specific primers for the mecA gene. The results showed that the Pi-Lise rapid extraction reagent was able to efficiently extract nucleic acids from the analyzed bacteria, maintaining the integrity of the genetic material and presenting similarity with the results obtained by more traditional extraction columns. This innovative approach demonstrates a promising alternative for the rapid extraction of nucleic acids from bacterial samples, becoming an indication of its increasing use in molecular techniques. The use of rapid nucleic acid extraction reagents can be considered a valuable strategy to accelerate and optimize molecular diagnostic procedures in microbiology, allowing greater speed and cost reduction. Palavras-chave: Molecular diagnosis, Nucleic acid extraction, PCR Agência de fomento:CNPq, CAPES, FAPEMIG | |||||