Automated method based in VNTR analysis for Rickettsiae genotyping.
A genetic locus named Rc-65, which is 5' adjacent to gene dksA and 3' adjacent to xerC gene, has previously been demonstrated to contain a VNTR with high discriminatory power in several rickettsial strains and thus, potentially useful for genetically similar strains identification. In this work, we present an automated molecular identification method based on capillary electrophoresis separation of VNTRs amplicons. The resulting electropherograms were in agreement with the sequence data obtained in a previous work. The presented genotyping method is fast and suitable for full automation, being a powerful tool for epidemiological surveillance in a large number of samples and enables the detection co-infected samples. The combination of other VNTR loci should improve the discriminatory capacity of this typing system, providing greater resolution and contributing to a more accurate VNTR-based assay. To our knowledge, this is the first automated assay for rickettsial strains identification.
Vitorino
Liliana
L
CEVDI, Instituto Nacional de Saudé Dr. Ricardo Jorge, INSA, Edifíco LEMES, Av. Padre Cruz, 1649-016 Lisboa, Portugal.
de Sousa
Rita
R
Bacellar
Fatima
F
Zé-Zé
Líbia
L
D015373:Bacterial Typing Techniques; D002874:Chromosome Mapping; D005838:Genotype; D018598:Minisatellite Repeats; D016133:Polymerase Chain Reaction / Q000379:methods; D012281:Rickettsia / Q000145:classification / Q000235:genetics*; D020114:Rickettsia conorii / Q000235:genetics*