DNA fingerprinting of pathogenic bacteria by fluorophore-enhanced repetitive sequence-based polymerase chain reaction.
Fluorophore-labeled oligonucleotide primers complementary to defined interspersed repetitive sequences conserved in diverse bacteria were used in the polymerase chain reaction to generate DNA fingerprint patterns from selected pathogenic bacteria. Fluorophore-enhanced, repetitive sequence-based polymerase chain reaction allowed discrimination between unrelated isolates of penicillin-resistant Streptococcus pneumoniae recovered from pediatric patients and Mycobacterium avium cultured from patients with acquired immunodeficiency syndrome. Combinations of oligonucleotide primers labeled with distinct fluorescent dyes enabled simultaneous DNA fingerprinting and Shiga-like toxin gene detection in enterohemorrhagic Escherichia coli isolates. Fluorophore-enhanced, repetitive sequence-based polymerase chain reaction was performed with either purified DNA or intact cells that were lysed during the polymerase chain reaction. Fluorophore-enhanced, repetitive sequence-based polymerase chain reaction successfully combines polymerase chain reaction amplification and fluorescent label detection for DNA fingerprinting of cultured bacterial pathogens.
Versalovic
J
J
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030.
Kapur
V
V
Koeuth
T
T
Mazurek
G H
GH
Whittam
T S
TS
Musser
J M
JM
Lupski
J R
JR
D001483:Base Sequence; D016172:DNA Fingerprinting / Q000379:methods*; D004926:Escherichia coli / Q000302:isolation & purification; D005453:Fluorescence; D008969:Molecular Sequence Data; D009162:Mycobacterium avium / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid; D013296:Streptococcus pneumoniae / Q000302:isolation & purification