Genomic fingerprinting of Bartonella species by repetitive element PCR for distinguishing species and isolates.
Repetitive-element PCR (rep-PCR) with primers based on repetitive extragenic palindromic (REP) and enterobacterial repetitive intergenic consensus (ERIC) repeated DNA sequences was used for genomic finger-printing of Bartonella species. This technique was applied by using either extracted genomic DNA or preparations of whole bacterial cells directly. PCR fingerprints with either the REP-based primers (REP-PCR) or primers based on the ERIC repeat (ERIC-PCR) revealed species-specific band patterns for the various Bartonella isolates. DNA fingerprints obtained from rep-PCR of extracted genomic DNA or from preparations of whole cells yielded comparable patterns. ERIC-PCR banding patterns were less complex than those obtained by REP-PCR but allowed better discrimination between strains within species. By combining results of REP-PCR and ERIC-PCR, five different fingerprint profiles were identified among 17 isolates of Bartonella henselae, but only one profile was identified among the five isolates of Bartonella quintana. Other Bartonella species yielded distinct rep-PCR fingerprints. rep-PCR is a useful technique for identification of Bartonella organisms to the species level and offers the advantage of ease of performance, with only small quantities of cells needed for the whole-cell procedure. This technique also appears to be useful for subtyping B. henselae isolates.
Rodriguez-Barradas
M C
MC
Section of Infectious Diseases, Medical Service, Veterans Affairs Medical Center, Houston, TX 77030, USA.
Hamill
R J
RJ
Houston
E D
ED
Georghiou
P R
PR
Clarridge
J E
JE
Regnery
R L
RL
Koehler
J E
JE
D017088:AIDS-Related Opportunistic Infections / Q000382:microbiology; D000818:Animals; D015373:Bacterial Typing Techniques; D001473:Bartonella / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D001474:Bartonella Infections / Q000382:microbiology; D002415:Cats; D016384:Consensus Sequence; D016172:DNA Fingerprinting / Q000379:methods*; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics; D005069:Evaluation Studies as Topic; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid; D013045:Species Specificity
D017931:DNA Primers; D004269:DNA, Bacterial