Automated laser fluorescence analysis of randomly amplified polymorphic DNA: a rapid method for investigating nosocomial transmission of Acinetobacter baumannii.
A rapid method for genotyping Acinetobacter baumannii based on PCR-fingerprinting with fluorescent primers was evaluated. Automated laser fluorescence analysis (ALFA) enabled on-line generation of high resolution DNA-fingerprints during polyacrylamide gel electrophoresis of randomly amplified polymorphic DNA (RAPD) products. The results were in concordance with macro-restriction fragment patterns produced by pulse-field gel electrophoresis (PFGE) of ApaI digests of chromosomal DNA. RAPD-ALFA was able to identify homologous strains suggestive of horizontal transmission in < 8 h after colonies were obtained on solid media, whereas PFGE analysis took c. 90 h. Speed and digitised data format renders RAPD-ALFA attractive for routine in-house epidemiological screening of isolates from intensive care and other hospital units.
D000150:Acinetobacter / Q000145:classification / Q000235:genetics*; D000151:Acinetobacter Infections / Q000453:epidemiology / Q000382:microbiology / Q000635:transmission*; D001483:Base Sequence; D002876:Chromosomes, Bacterial / Q000235:genetics; D003428:Cross Infection / Q000453:epidemiology / Q000382:microbiology / Q000635:transmission*; D016172:DNA Fingerprinting / Q000379:methods; D017931:DNA Primers / Q000737:chemistry; D004269:DNA, Bacterial / Q000032:analysis*; D004196:Disease Outbreaks; D016521:Electrophoresis, Gel, Pulsed-Field; D004591:Electrophoresis, Polyacrylamide Gel; D005453:Fluorescence; D005456:Fluorescent Dyes; D005784:Gene Amplification / Q000235:genetics*; D005838:Genotype; D005858:Germany / Q000453:epidemiology; D006801:Humans; D007834:Lasers; D008969:Molecular Sequence Data
D017931:DNA Primers; D004269:DNA, Bacterial; D005456:Fluorescent Dyes