TÉCNICA DE PCR SIMPLEX ASSOCIADA A MÉTODO DE EXTRAÇÃO DE DNA DE BAIXO CUSTO PARA IDENTIFICAÇÃO MOLECULAR DE ISOLADOS CLÍNICOS DE SHIGELLA SPP 1 THE PCR SIMPLEX TECHNIQUE ASSOCIATED TO THE LOW COST DNA EXTRACTION METHOD FOR MOLECULAR IDENTIFICATION OF SHIGELLA SPP CLINICAL ISOLATES

Shigellosis is an enteric disease responsible for 1.1 million deaths annually. 60% of those is represented by children under 5 years old. Shigella species are found in contaminated food and water, so the transmissions route is through fecal-oral contact. The present study aims to adapt a molecular method for the identification of clinical gastroenteritis cases caused by Shigella spp. The bacterial culture of the S. sonnei LCL-01 clinical isolate was subjected to the genomic DNA extraction by means of the modified phenol-chloroform method. The resulting genetic material was subjected to the PCR simplex technique. This procedure resulted in a genomic DNA of good quality, with a fragment of 615 pb after the amplification of the ipaH gene. In this manner, it was possible the molecular identification of S.sonnei by adapting an in-house low cost method, which is based in the usage of phenol-chloroform as a tool for breaking the bacteria cell wall for the extraction of genomic DNA. This may help tracking, detecting and treating infections caused by S. sonnei.