Multicenter evaluation of arbitrarily primed PCR for typing of Staphylococcus aureus strains.
Fifty-nine isolates of Staphylococcus aureus and a single strain of Staphylococcus intermedius were typed by arbitrarily primed PCR (AP-PCR). To study reproducibility and discriminatory abilities, AP-PCR was carried out in seven laboratories with a standardized amplification protocol, template DNA isolated in a single institution, and a common set of three primers with different resolving powers. The 60 strains could be divided into 16 to 30 different genetic types, depending on the laboratory. This difference in resolution was due to differences in technical procedures (as shown by the deliberate introduction of experimental variables) and/or the interpretation of the DNA fingerprints. However, this did not hamper the epidemiologically correct clustering of related strains. The average number of different genotypes identified exceeded those of the more traditional typing strategies (F. C. Tenover, R. Arbeit, G. Archer, J. Biddle, S. Byrne, R. Goering, G. Hancock, G. A. Hebert, B. Hill, R. Hollis, W. R. Jarvis, B. Kreiswirth, W. Eisner, J. Maslow, L. K. McDougal, J. M. Miller, M. Mulligan, and M. A. Pfaller, J. Clin. Microbiol. 32:407-415, 1994). Comparison of AP-PCR with pulsed-field gel electrophoresis (PFGE) indicated the existence of strains with constant PFGE types but variable AP-PCR types. The reverse (constant AP-PCR and variable PFGE patterns) was also observed. This indicates additional resolution for combined analyses. It is concluded that AP-PCR is well suited for genetic analysis and monitoring of nosocomial spreading of staphylococci. The interlaboratory reproducibility of DNA-banding patterns and the intralaboratory standardization need improvement.
van Belkum
A
A
Department of Bacteriology, University Hospital Dijkzigt, Rotterdam, The Netherlands.
Kluytmans
J
J
van Leeuwen
W
W
Bax
R
R
Quint
W
W
Peters
E
E
Fluit
A
A
Vandenbroucke-Grauls
C
C
van den Brule
A
A
Koeleman
H
H
D015373:Bacterial Typing Techniques / Q000706:statistics & numerical data; D001483:Base Sequence; D016172:DNA Fingerprinting; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004196:Disease Outbreaks; D016521:Electrophoresis, Gel, Pulsed-Field; D005069:Evaluation Studies as Topic; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D015203:Reproducibility of Results; D013045:Species Specificity; D013203:Staphylococcal Infections / Q000453:epidemiology / Q000382:microbiology; D013211:Staphylococcus aureus / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification
D017931:DNA Primers; D004269:DNA, Bacterial