Usefulness of a multiplex PCR for the rapid identification of Candida glabrata species complex in Mexican clinical isolates
Por:
Reyes-Montes M.R., Acosta-Altamirano, Gustavo, Duarte-Escalante, Esperanza, Salazar E.G., Martinez-Herrera, Erick, Arenas, Roberto, Gonzalez, Gloria, Frías-De-León M.G.
Publicada:
1 ene 2019
Resumen:
Candida glabrata complex includes three species identified through
molecular biology methods: C. glabrata sensu stricto, C. nivariensis and
C. bracarensis. In Mexico, the phenotypic methods are still used in the
diagnosis; therefore, the presence of C. nivariensis and C. bracarensis
among clinical isolates is still unknown. The aim of this study was to
evaluate the utility of a multiplex PCR for the identification of the C.
glabrata species complex. DNA samples from 92 clinical isolates that
were previously identified through phenotypic characteristics as C
glabrata were amplified by four oligonucleotides (UNI-5.8S, GLA-f,
BRA-f, and NIV-f) that generate amplicons of 397, 293 and 223-bp
corresponding to C. glabrata sensu stricto, C. nivariensis. and C.
bracarensis, respectively. The amplicon sequences were used to perform a
phylogenetic analysis through the Maximum Likelihood method (MEGA6),
including strains and reference sequences of species belonging to C.
glabrata complex. In addition, recombination and linkage disequilibrium
were estimated (DnaSP version 5.0) for C. glabrata sensu stricto
isolates. Eighty-eight isolates generated a 397-bp fragment and only in
one isolate a 223-bp amplicon was observed. In the phylogenetic tree,
the sequences of 397-bp were grouped with C. glabrata reference
sequences, and the sequence of 223-bp was grouped with C. bracarensis
reference sequences, corroborating the PCR identification. The number of
recombination events for the isolates of C. glabrata sensu stricto was
zero, suggesting a clonal population structure. Three isolates that did
not amplify any of the expected fragments were identified as
Saccharomyces cerevisiae through the sequencing of the D1/D2 domain
region within the 28S rDNA gene. The multiplex PCR is a fast,
cost-effective and reliable tool that can be used in clinical
laboratories to identify C. glabrata complex species.
Filiaciones:
Reyes-Montes M.R.:
Universidad Nacional Autónoma de México, Facultad de Medicina, Departamento de Microbiología y Parasitología, Ciudad de México, Mexico
Acosta-Altamirano, Gustavo:
Hospital Regional de Alta Especialidad de Ixtapaluca, Unidad de Investigación, Ixtapaluca, Mexico
Hosp Reg Alta Especialidad Ixtapaluca, Ixtapaluca, Mexico
Duarte-Escalante, Esperanza:
Universidad Nacional Autónoma de México, Facultad de Medicina, Departamento de Microbiología y Parasitología, Ciudad de México, Mexico
Univ Nacl Autonoma Mexico, Dept Microbiol & Parasitol, Ciudad De Mexico, Mexico
Salazar E.G.:
Hospital Regional de Alta Especialidad de Ixtapaluca, Unidad de Investigación, Ixtapaluca, Mexico
Martinez-Herrera, Erick:
Hospital Regional de Alta Especialidad de Ixtapaluca, Unidad de Investigación, Ixtapaluca, Mexico
Hosp Reg Alta Especialidad Ixtapaluca, Ixtapaluca, Mexico
Arenas, Roberto:
Hospital General “Dr. Manuel Gea González, Sección de Micología, Ciudad de México, Mexico
Hosp Gen Dr Manuel Gea Gonzalez, Secc Micol, Ciudad De Mexico, Mexico
Gonzalez, Gloria:
Universidad Autónoma de Nuevo León, Facultad de Medicina, Departamento de Microbiología, Monterrey, Mexico
Univ Autonoma Nuevo Leon, Fac Med, Dept Microbiol, Monterrey, Mexico
Frías-De-León M.G.:
Hospital Regional de Alta Especialidad de Ixtapaluca, Unidad de Investigación, Ixtapaluca, Mexico
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