Whole-cell repetitive element sequence-based polymerase chain reaction allows rapid assessment of clonal relationships of bacterial isolates.
Repetitive element sequence-based polymerase chain reaction (rep-PCR) enables the generation of DNA fingerprints which discriminate bacterial species and strains. We describe the application of whole-cell methods which allow specimens from broth cultures or colonies from agar plates to be utilized directly in the PCR mixture. The rep-PCR-generated DNA fingerprints obtained with whole-cell samples match results obtained with genomic DNA templates. Examples with different gram-negative bacteria (e.g., Citrobacter diversus, Escherichia coli, and Pseudomonas aeruginosa) and gram-positive bacteria (e.g., Staphylococcus aureus and Streptococcus pneumoniae) are demonstrated. Rapid specimen preparation methods enable rep-PCR-based fingerprinting to be completed in several hours and, therefore, allows the timely analysis of epidemiological relationships.
Woods
C R
CR
Department of Pediatrics, Baylor College of Medicine, Houston, Texas 77030.
Versalovic
J
J
Koeuth
T
T
Lupski
J R
JR
D001431:Bacteriological Techniques; D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000235:genetics; D005069:Evaluation Studies as Topic; D006090:Gram-Negative Bacteria / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D006094:Gram-Positive Bacteria / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid
D004269:DNA, Bacterial