Patent ID: 6664080
Filing Date: 2003-12-16
Classification: C12Q

Abstract:
A method for detection and differentiation of pathogenic enterobacteria in a sample, said method comprising:isolating nucleic acid from said sample; adding a set of oligonucleotide primer pairs to said nucleic acid sample wherein said set of oligonucleotide primers comprises at least five oligonucleotide primer pairs wherein, wherein each primer pair specifically amplifies a DNA sequence of a virulence factor/toxin gene characteristic for one each of the subgroups of the pathogenic E. coli strains, said subgroups comprising enterotoxigenic, enteroaggregative, enteroinvasive, enteropathogenic and enterohemorrhagic E. coli strains and wherein for amplification of each subgroup at least one oligonucleotide primer pair is included in said set of oligonucleotide primer pairs, and wherein the set comprises a primer pair that hybridizes to an inv-plasmid of enteroinvasive E. coli and wherein said primer pair is El-1: 5â€³TTT CTG GAT GGT ATG GTG AGG 3â€² (SEQ ID NO: 9) and E1-2: 5â€²CTT GAA CAT AAG GAA ATA AAC 3â€² (SEQ ID NO: 10) which specifically amplifies a DNA sequence of an inv plasmid characteristic for enteroinvasive E. coli; and subjecting said nucleic acid and said set of primer pairs to an amplification process; and detecting the presence of at least one amplified product, wherein the presence of at least one amplified product indicates the presence of at least one pathogenic enterobactcria strain in said sample.