DNA-FINGERPRINTING OF PATHOGENIC BACTERIA BY FLUOROPHORE-ENHANCED REPETITIVE SEQUENCE-BASED POLYMERASE CHAIN-REACTION

Fluorophore-labeled oligonucleotide primers complementary to defined i nterspersed repetitive sequences conserved in diverse bacteria were us ed in the polymerase chain reaction to generate DNA fingerprint patter ns from selected pathogenic bacteria. Fluorophore-enhanced, repetitive sequence-based polymerase chain reaction allowed discrimination betwe en unrelated isolates of penicillin-resistant Streptococcus pneumoniae recovered from pediatric patients and Mycobacterium avium cultured fr om patients with acquired immunodeficiency syndrome. Combinations of o ligonucleotide primers labeled with distinct fluorescent dyes enabled simultaneous DNA fingerprinting and Shiga-like toxin gene detection in enterohemorrhagic Escherichia coli isolates. Fluorophore-enhanced, re petitive sequence-based polymerase chain reaction was performed with e ither purified DNA or intact cells that were lysed during the polymera se chain reaction. Fluorophore-enhanced, repetitive sequence-based pol ymerase chain reaction successfully combines polymerase chain reaction amplification and fluorescent label detection for DNA fingerprinting of cultured bacterial pathogens.