Evaluation of a highly discriminating multiplex multi-locus variable-number of tandem-repeats (MLVA) analysis for Vibrio cholerae.
Vibrio cholerae is the etiological agent of cholera and may be used in bioterror actions due to the easiness of its dissemination, and the public fear for acquiring the cholera disease. A simple and highly discriminating method for connecting clinical and environmental isolates of V. cholerae is needed in microbial forensics. Twelve different loci containing variable numbers of tandem-repeats (VNTRs) were evaluated in which six loci were polymorphic. Two multiplex reactions containing PCR primers targeting these six VNTRs resulted in successful DNA amplification of 142 various environmental and clinical V. cholerae isolates. The genetic distribution inside the V. cholerae strain collection was used to evaluate the discriminating power (Simpsons Diversity Index=0.99) of this new MLVA analysis, showing that the assay have a potential to differentiate between various strains, but also to identify those isolates which are collected from a common V. cholerae outbreak. This work has established a rapid and highly discriminating MLVA assay useful for track back analyses and/or forensic studies of V. cholerae infections.
Olsen
Jaran S
JS
Norwegian Defence Research Establishment, P. O. Box 25, N-2027 Kjeller, Norway. Jaran-Strand.Olsen@ffi.no
Aarskaug
Tone
T
Skogan
Gunnar
G
Fykse
Else Marie
EM
Ellingsen
Anette Bauer
AB
Blatny
Janet M
JM
D015373:Bacterial Typing Techniques / Q000379:methods*; D002771:Cholera / Q000175:diagnosis / Q000382:microbiology; D016172:DNA Fingerprinting / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics*; D004783:Environmental Microbiology; D005838:Genotype; D006801:Humans; D018598:Minisatellite Repeats; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D012680:Sensitivity and Specificity; D014734:Vibrio cholerae / Q000145:classification* / Q000235:genetics*
D017931:DNA Primers; D004269:DNA, Bacterial