Disparity between Multilocus Enzyme Electrophoresis, Microsatellite Markers and Pulsed-Field Gel Electrophoresis in epidemiological tracking of Candida albicans |
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| Authors: | Marcelo Fabiano Gomes Boriollo Ricardo Antunes Dias Nelma de Mello Silva Oliveira Henrique Marques Barbosa de Souza Aline Aparecida Pizzirani-Kleiner |
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| Institution: | a Laboratory of Molecular Biology and Genetics, Faculty of Medical Sciences, University of Alfenas, Minas Gerais, Brazilb Laboratory of Biology and Physiology of Microorganisms, Faculty of Medical Sciences, University of Alfenas, Minas Gerais, Brazilc Laboratory of Oral Microbiology, Department of Oral Pathology and Physiology, Faculty of Ordontology of Araraquara, State University of São Paulo, Araraquara, São Paulo, Brazild Laboratory of Plant Improvement, Center of Nuclear Energy in Agriculture, University of São Paulo, Piracicaba, SP, Brazile Department of Genetics, Escola Superior de Agricultura Luiz de Queiroz, University of São Paulo, Piracicaba, SP, Brazil |
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| Abstract: | Various molecular systems are available for epidemiological, genetic, evolutionary, taxonomic and systematic studies of innumerable fungal infections, especially those caused by the opportunistic pathogen C. albicans. A total of 75 independent oral isolates were selected in order to compare Multilocus Enzyme Electrophoresis (MLEE), Electrophoretic Karyotyping (EK) and Microsatellite Markers (Simple Sequence Repeats - SSRs), in their abilities to differentiate and group C. albicans isolates (discriminatory power), and also, to evaluate the concordance and similarity of the groups of strains determined by cluster analysis for each fingerprinting method. Isoenzyme typing was performed using eleven enzyme systems: Adh, Sdh, M1p, Mdh, Idh, Gdh, G6pdh, Asd, Cat, Po, and Lap (data previously published). The EK method consisted of chromosomal DNA separation by pulsed-field gel electrophoresis using a CHEF system. The microsatellite markers were investigated by PCR using three polymorphic loci: EF3, CDC3, and HIS3. Dendrograms were generated by the SAHN method and UPGMA algorithm based on similarity matrices (SSM). The discriminatory power of the three methods was over 95%, however a paired analysis among them showed a parity of 19.7-22.4% in the identification of strains. Weak correlation was also observed among the genetic similarity matrices (SSMMLEE × SSMEK × SSMSSRs). Clustering analyses showed a mean of 9 ± 12.4 isolates per cluster (3.8 ± 8 isolates/taxon) for MLEE, 6.2 ± 4.9 isolates per cluster (4 ± 4.5 isolates/taxon) for SSRs, and 4.1 ± 2.3 isolates per cluster (2.6 ± 2.3 isolates/taxon) for EK. A total of 45 (13%), 39 (11.2%), 5 (1.4%) and 3 (0.9%) clusters pairs from 347 showed similarity (SJ) of 0.1-10%, 10.1-20%, 20.1-30% and 30.1-40%, respectively. Clinical and molecular epidemiological correlation involving the opportunistic pathogen C. albicans may be attributed dependently of each method of genotyping (i.e., MLEE, EK, and SSRs) supplemented with similarity and grouping analysis. Therefore, the use of genotyping systems that give results which offer minimum disparity, or the combination of the results of these systems, can provide greater security and consistency in the determination of strains and their genetic relationships. |
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| Keywords: | Candida albicans Cluster Analysis Electrophoretic Karyotyping Epidemiological Tracking Microsatellite Markers Multilocus Enzyme Electrophoresis |
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