Selection of Polymorphic Patterns Obtained by RAPD-PCR through Qualitative and Quantitative Analyses to Differentiate Aspergillus fumigatus
Por:
Valencia-Ledezma, Omar E., Castro-Fuentes, Carlos A., Duarte-Escalante, Esperanza, Guadalupe Frias-De-Leon, Maria, del Rocio Reyes-Montes, Maria
Publicada:
1 mar 2022
Resumen:
The objective of this work was to use the random amplification of the
polymorphic DNA-polymerase chain reaction (RAPD-PCR) technique to select
polymorphic patterns through qualitative and quantitative analyses to
differentiate the species A. flavus, A. fumigatus, A. niger and A.
tubingensis. Twenty-seven Aspergillus isolates from different species
were typified using phenotypic (macro- and micromorphology) and
genotypic (partial BenA gene sequencing) methods. Thirty-four primers
were used to obtain polymorphic patterns, and with these a qualitative
analysis was performed to select the primers that presented
species-specific patterns to distinguish each species. For the
quantitative selection, a database was built from the polymorphic
patterns and used for the construction of logistic regression models;
later, the model that presented the highest value of sensitivity against
specificity was evaluated through ROC curves. The qualitative selection
showed that the primers OPA-19, P54, 1253 and OPA-02 could differentiate
the species. A quantitative analysis was carried out through logistic
regression, whereby a species-specific correlation of sensitivity and
specificity greater than 90% was obtained for the primers: OPC-06 with
a 96.32% match to A. flavus; OPF-01 with a 100% match to A. fumigatus;
OPG-13 with a 98.01% match to A. tubingensis; and OPF-07 with a 99.71%
match to A. niger. The primer OPF-01 discriminated the four species as
well as closely related species. The quantitative methods using the
selected primers allowed discrimination between species and showed their
usefulness for genotyping some of the species of medical relevance
belonging to the genus Aspergillus.
Filiaciones:
Valencia-Ledezma, Omar E.:
Univ Nacl Autonoma Mexico, Fac Med, Dept Microbiol & Parasitol, Unidad Micol, Mexico City 04510, DF, Mexico
Castro-Fuentes, Carlos A.:
Univ Nacl Autonoma Mexico, Fac Med, Dept Microbiol & Parasitol, Unidad Micol, Mexico City 04510, DF, Mexico
Duarte-Escalante, Esperanza:
Univ Nacl Autonoma Mexico, Fac Med, Dept Microbiol & Parasitol, Unidad Micol, Mexico City 04510, DF, Mexico
Guadalupe Frias-De-Leon, Maria:
Hosp Reg Alta Especialidad Ixtapaluca, Carretera Fed Mexico Puebla Km 34-5, Pueblo De Zoquiapan 56530, Ixtapaluca, Mexico
del Rocio Reyes-Montes, Maria:
Univ Nacl Autonoma Mexico, Fac Med, Dept Microbiol & Parasitol, Unidad Micol, Mexico City 04510, DF, Mexico
Green Published, gold, All Open Access, Gold, Green
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