Optimal DNA isolation method for detection of bacteria in clinical specimens by using the technique of PCR

Background and objective: enteropathogenic escherichia coli is an important cause of diarrhea in infants,especially in regions of poor sanitation. present study tried to detect enteropathogenic e.coli and its gene bfp in stool samples of diarrheal children by polymerase chain reaction in order to facilitate the diagnosis. patients and method: forty diarrheal sample of children age range 1-3 years were collected from different clinical laboratories in lahore in a period of two-three months. polymerase chain reaction of dna was carried out by using taq dna polymerase,and template dna. results: out of 40 patients' samples,5 samples were detected as pcr positive for bfp,when specific primer for bfp gene was used resulting in amplification of 330 base pair segment. the rest of samples were negative in bpf amplification. identification of e.coli and its dna was carried out. conclusion:it is therefore concluded that broad-range amplification of bacterial dna from complicated cases has proved useful for the diagnosis of e.coli.