URA5-RFLP is one of the most widely used genotyping methods relating to Cryptococcus neoformans and C. gattii consensus genotype nomenclature. In order to identify a molecular type, this method uses a visual comparison of digested PCR products of tested and reference strains, therefore any anomaly in RFLP patterns of studied isolates makes recognition difficult or impossible. This report describes a strain of VNIV type showing an atypical URA5-RFLP pattern as well as a group of AD hybrids displaying the same anomaly. The atypical RFLP pattern is the result of a point mutation and emergence of a new restriction site. Emergence of the allele presenting a new banding pattern may lead to misidentification using the URA5-RFLP technique; the results of this study as well as the literature data may suggest the spread of the allele in the environment.

• MeSH
• Cryptococcus neoformans klasifikace   genetika   MeSH
• genotyp MeSH
• geny hub genetika   MeSH
• mikrobiologie životního prostředí MeSH
• mutace MeSH
• mykologické určovací techniky MeSH
• orotátfosforibosyltransferasa genetika   MeSH
• polymorfismus délky restrikčních fragmentů MeSH
• sekvence nukleotidů MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• orotátfosforibosyltransferasa MeSH

Cryptococcosis is caused by members of the Cryptococcus neoformans/Cryptococcus gattii species complex. Based on molecular identification, these two species have been further differentiated into molecular types. The aim of this work was to characterize clinical cryptococcal isolates recovered from six hospitals in Northeast Mexico from 1995 to 2011. One hundred and sixty-six isolates, which were characterized by biochemical tests and in vitro susceptibility to amphotericin B, fluconazole, and voriconazole, and M13 PCR fingerprinting, were included in this study. Utilizing phenotypic tests, 153 isolates (92.16 %) were identified as C. neoformans and 13 (7.83 %) as C. gattii. All isolates were susceptible to all antifungals tested. Employing M13 PCR fingerprinting, eight molecular types were detected. VNI was the most common genotype (124 cases; 74.6 %), followed by VNII (15 cases; 9 %), VNIII (8 cases; 4.8 %), VNIV (6 cases; 3.6 %), VGI (6 cases; 3.6 %), VGII (3 cases; 1.8 %), and VGIII and VGIV (2 cases, 1.2 % each). We confirm the presence of C. gattii in clinical isolates in Northeast Mexico, and a high clonal diversity in the studied strains of C. neoformans/C. gattii species complex.

• MeSH
• antifungální látky farmakologie   MeSH
• Cryptococcus gattii klasifikace   genetika   izolace a purifikace   MeSH
• Cryptococcus neoformans klasifikace   genetika   izolace a purifikace   MeSH
• DNA fingerprinting MeSH
• dospělí MeSH
• kryptokokóza epidemiologie   mikrobiologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• mladiství MeSH
• mladý dospělý MeSH
• molekulární epidemiologie MeSH
• molekulární typizace * MeSH
• mykologické určovací techniky * MeSH
• nemocnice MeSH
• polymerázová řetězová reakce MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Mexiko epidemiologie   MeSH
• Názvy látek
• antifungální látky MeSH

The cryptococcal polysaccharide antigen was detected in 10 cerebrospinal fluid (CSF) and 23 serum samples from cryptococcal meningitis and intestinal cryptococcosis by the cryptococcal antigen latex agglutination system (CALAS). CALAS titers in CSF and serum samples of cryptococcal meningitis ranged over 8-2048 and 32-2048, respectively, while in cases of intestinal cryptococcosis, serum titers ranged over 8-2048. The isolates of yeast Cryptococcus neoformans were determined to be of serotype A or of the A/D pair. The total leukocyte count and biochemical parameters in CSF were significantly increased as indicators of microbial infection. Furthermore, the in vitro change of the teleomorph (sexual state) to the anamorph (asexual state) was also detected and the teleomorph state changed in vivo to the encapsulated anamoph state which is more virulent during infection in vivo than the yeast-like noncapsulated form. Two primers for internal transcribed spacer (ITS) regions of ribosomal DNA were used for molecular detection of C. neoformans. After PCR amplification, a DNA band of 415 bp, visualized on agarose gel, indicated the presence of C. neoformans cells in the tested CSF and serum samples. The primer sensitivity was also characterized using purified yeast chromosomal DNA as template; it was about 20 pg or more chromosomal DNA which represents about 10 cells of C. neoformans. The primers were also specific for ITS regions of C. neoformans and gave negative results with Candida albicans and E. coli chromosomal DNA templates.

• MeSH
• Cryptococcus neoformans klasifikace   genetika   izolace a purifikace   MeSH
• dítě MeSH
• DNA fungální krev   mozkomíšní mok   MeSH
• dospělí MeSH
• kryptokoková meningitida diagnóza   mikrobiologie   MeSH
• kryptokokóza diagnóza   mikrobiologie   MeSH
• latex fixační testy MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mykologické určovací techniky MeSH
• nemoci střev diagnóza   mikrobiologie   MeSH
• polymerázová řetězová reakce metody   MeSH
• polysacharidy krev   mozkomíšní mok   MeSH
• předškolní dítě MeSH
• senzitivita a specificita MeSH
• sérotypizace MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• předškolní dítě MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• Geografické názvy
• Egypt MeSH
• Názvy látek
• cryptococcal polysaccharide MeSH Prohlížeč
• DNA fungální MeSH
• polysacharidy MeSH