TY - JOUR
T1 - Comparative usefulness of ribotyping, exotoxin A genotyping, and SalI restriction fragment length polymorphism analysis for Pseudomonas aeruginosa lineage assessment
AU - Nociari, Marcelo M.
AU - Catalano, Mariana
AU - García, Daniela Centrón
AU - Copenhaver, Steven C.
AU - Vasil, Michael L.
AU - Sordelli, Daniel O.
N1 - Funding Information:
Jaime Kovensky, Ana Di Martino, Alicia Rossi, Liliana Fio- rile,C arlos Bantar, and Lina Famiglietti for contributing bacterial strains and isolates. This study was supported in part by Consejo National de Investigaciones Cientificas y Tecnicas (CONICET), Buenos Aires, and by UBACYT, Universitdad
PY - 1996/4
Y1 - 1996/4
N2 - Ribotyping, exotoxin A genotyping (EAGP), and restriction fragment length polymorphism (RFLP) analysis of total DNA with Sail (Sail RFLP) were compared for intraspecies discrimination of 93 Pseudomonas aeruginosa isolates. Typeability of all methods was 100% and the results of typing with each method remained unchanged during laboratory manipulation. Clonal groups defined with each molecular method were largely coincident and, in those cases where inconsistencies were detected, isolates were analyzed by transverse alternating field gel electrophoresis (TAFE) and arbitrarily primed polymerase chain reaction (AP- PCR). SalI RFLP analysis was highly discriminative so as to distinguish unrelated isolates of close lineage. However, it was not a good method to identify isolates of unrelated lineage because SalI RFLP appeared to be subjected to convergent evolution. The index of discrimination suggested by Hunter and Gaston was determined to assess the discriminatory power of the molecular methods utilized either alone or in several combinations. Combined use of ribotyping and SalI RFLP analysis reached the highest index of discrimination (0.982) and proved to be a very valuable tool for epidemiological differentiation of P. aeruginosa isolates.
AB - Ribotyping, exotoxin A genotyping (EAGP), and restriction fragment length polymorphism (RFLP) analysis of total DNA with Sail (Sail RFLP) were compared for intraspecies discrimination of 93 Pseudomonas aeruginosa isolates. Typeability of all methods was 100% and the results of typing with each method remained unchanged during laboratory manipulation. Clonal groups defined with each molecular method were largely coincident and, in those cases where inconsistencies were detected, isolates were analyzed by transverse alternating field gel electrophoresis (TAFE) and arbitrarily primed polymerase chain reaction (AP- PCR). SalI RFLP analysis was highly discriminative so as to distinguish unrelated isolates of close lineage. However, it was not a good method to identify isolates of unrelated lineage because SalI RFLP appeared to be subjected to convergent evolution. The index of discrimination suggested by Hunter and Gaston was determined to assess the discriminatory power of the molecular methods utilized either alone or in several combinations. Combined use of ribotyping and SalI RFLP analysis reached the highest index of discrimination (0.982) and proved to be a very valuable tool for epidemiological differentiation of P. aeruginosa isolates.
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U2 - 10.1016/0732-8893(96)00029-6
DO - 10.1016/0732-8893(96)00029-6
M3 - Article
C2 - 8831031
AN - SCOPUS:0030130996
SN - 0732-8893
VL - 24
SP - 179
EP - 190
JO - Diagnostic Microbiology and Infectious Disease
JF - Diagnostic Microbiology and Infectious Disease
IS - 4
ER -