Typing of Listeria monocytogenes by random amplified polymorphic DNA (RAPD) analysis.
The aim of the study was to determine the effectiveness of random amplified polymorphic DNA analysis in typing Listeria monocytogenes from human infections. Twenty-five L. monocytogenes serogroup 1/2 and 70 serogroup 4 including 14 serovar 4b(x) were typed by RAPD-PCR analysis. Six primers were used to type each L. monocytogenes isolate and the DNA amplification performed with supertaq DNA polymerase in a Hybaid Thermal Reactor. Each bacterial strain was analysed separately with all primers and the profiles were judged by eye and designated to a group by comparison to other strains. Bands were classified as major or minor. Based on analysis of major band patterns, the 25 serogroup 1/2 isolates gave rise to 12 different groups. The groups only contained serovar 1/2a or 1/2b with a single exception. Using minor bands all isolates could be distinguished. All the serogroup 4 isolates gave the same major band patterns. The 14 serovar 4b(x) isolates which were epidemiologically related gave identical profiles with the exception of one isolate. Of the remaining strains, 41 produced individual patterns on minor band analysis. RAPD analysis with multiple primers is low cost, discriminatory and is most ideally suited to testing small (< 50) numbers of strains. We have shown that serogroup 1/2 L. monocytogenes strains are a more diverse group than serovar 4b strains and RAPD-PCR will provide a technique of considerable value in typing L. monocytogenes in the future.
O'Donoghue
K
K
Department of Medical Microbiology, Southmead Health Services NHS Trust, Westbury-on-Trym, Bristol, UK.
Bowker
K
K
McLauchlin
J
J
Reeves
D S
DS
Bennett
P M
PM
MacGowan
A P
AP
D001434:Bacteriophage Typing; D001483:Base Sequence; D003362:Cost-Benefit Analysis; D017931:DNA Primers; D006801:Humans; D008089:Listeria monocytogenes / Q000145:classification*; D008088:Listeriosis / Q000382:microbiology*; D008969:Molecular Sequence Data; D019105:Random Amplified Polymorphic DNA Technique
D017931:DNA Primers