25º Congresso Brasileiro de Microbiologia

25º Congresso Brasileiro de Microbiologia

ResumoID:2013-1

Área: Microbiologia Clinica ( Divisão A )

EVALUATION OF FIVE DIFFERENT METHODS TO EXTRACT MICROBIAL GENOMIC DNA FROM STOOL SAMPLES

Ila Fernanda Nunes Lima (UFC); Josiane da Silva Quetz (UFC); Alexandre Havt (UFC); Aldo Ângelo Moreira Lima (UFC)

Resumo

INTRODUCTION: Intestinal pathogens are usually diagnosed by isolation and identification of the organisms from fecal cultures. Culture methods are precise, but relatively time-consuming, labor-intensive and difficult for some fastidious intestinal pathogens. Polymerase chain reaction (PCR) is efficient, specific and a sensitive technique for the detection of organisms and are increasingly applied to the diagnosis of infectious diseases. We aimed to evaluate five different methods to extract DNA from stool samples to be used as a PCR template.

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RESULTS: Nested PCR amplification of the 18S rRNA gene of Cryptosporidium sp. and a single PCR of the aggR gene of EAEC showed differences among the types of pre-treatment tested. Analysis of gel bands intensity revealed variations among the methods, although all of them had originated bands clearly visible. For Cryptosporidium sp., the techniques 1 (physical + chemical pretreatment), 2 (chemical pretreatment only), and 5 (no pretreatment) resulted bands with similar intensity. For EAEC, the option 5 yielded the best level of detection.

FINANCIAL SUPPORT: FIC/NIH (USA) and CNPq (Brazil)

Palavras-chave: DIAGNÓSTICO MOLECULAR, DIARRÉIA INFANTIL, EXTRAÇÃO DE DNA

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