Identification and DNA fingerprinting of Legionella strains by randomly amplified polymorphic DNA analysis.
The randomly amplified polymorphic DNA (RAPD) technique was used in the development of a fingerprinting (typing) and identification protocol for Legionella strains. Twenty decamer random oligonucleotide primers were screened for their discriminatory abilities. Two candidate primers were selected. By using a combination of these primers, RAPD analysis allowed for the differentiation between all different species, between the serogroups, and further differentiation between subtypes of the same serogroup. The usefulness of RAPD analysis was also evaluated with outbreak-related clinical and environmental isolates previously typed by the restriction fragment length polymorphism technique. RAPD analysis proved to be as accurate as other genotypic methods, reproducible, and highly discriminatory and is a valuable new alternative to traditional fingerprinting and identification of Legionella species and strains.
Bansal
N S
NS
Division of Analytical Laboratories, Western Sydney Area Health Service, Lidcombe, New South Wales, Australia.
McDonell
F
F
D000818:Animals; D016172:DNA Fingerprinting / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis; D004196:Disease Outbreaks; D004783:Environmental Microbiology; D006801:Humans; D007875:Legionella / Q000145:classification* / Q000302:isolation & purification*; D007876:Legionellosis / Q000175:diagnosis / Q000453:epidemiology; D017720:Molecular Epidemiology; D012150:Polymorphism, Restriction Fragment Length; D019105:Random Amplified Polymorphic DNA Technique; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004269:DNA, Bacterial