Multiple-locus variable-number tandem repeat analysis of Legionella pneumophila using multi-colored capillary electrophoresis.
Several methods for typing of Legionella pneumophila exist, one of which is an 8-locus variable-number of tandem repeats analysis (MLVA). This method is based on separating and sizing amplified VNTR PCR products by agarose gel electrophoresis. In the present work, the existing L. pneumophila MLVA-8 assay is adapted to capillary electrophoresis. The assay was multiplexed by using multiple fluorescent labeling dyes and tested on a panel of L. pneumophila strains with known genotypes. The results from the capillary electrophoresis-based assay are shown to be equivalent to, and in a few cases more sensitive than, the gel-based genotyping assay. The assay presented here allows for a swift, automated and precise typing of L. pneumophila from patient or environmental samples and represents an improvement over the current gel-based method.
Nederbragt
Alexander J
AJ
University of Oslo, Department of Biology, Centre for Ecological and Evolutionary Synthesis, P.O. Box 1066 Blindern, N-0316 Oslo, Norway.
Balasingham
Anusha
A
Sirevåg
Reidun
R
Utkilen
Hans
H
Jakobsen
Kjetill S
KS
Anderson-Glenna
Mary J
MJ
D015373:Bacterial Typing Techniques / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005453:Fluorescence; D005838:Genotype; D016952:Legionella pneumophila / Q000145:classification* / Q000235:genetics*; D018598:Minisatellite Repeats; D017720:Molecular Epidemiology / Q000379:methods*; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods