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Laura Theill Catiana Dudiuk Soraya Morales-Lopez Indira Berrio José Yesid Rodríguez Adriana Marin Soledad Gamarra Guillermo Garcia-Effron 《Revista iberoamericana de micología》2018,35(2):110-112
Background
Candida auris and Candida haemulonii are emerging and multiresistant pathogens. C. auris has produced hospital outbreaks and is misidentified by phenotypic-based methods. The only reliable identification methods are DNA sequencing and MALDI-TOF.Aims
To develop a classical-PCR method capable of rapidly and accurately identify C. auris and C. haemulonii.Methods
A multiplex PCR was carried out in one tube that included an internal control and oligonucleotides that specifically hybridize to the ITS2 region of C. auris and C. haemulonii. The usefulness of the new method was verified by testing a collection of 50 strains of 20 different species (previously identified by ITS sequencing). The selection of species was made in order to emulate the C. auris panel used by the CDC to validate diagnostic tools. In addition, other yeast species not included in the aforementioned panel were incorporated based on reported identification errors.Results
The results obtained with the proposed protocol were in total agreement with those obtained by ITS sequencing.Conclusions
We present a PCR method able to unequivocally identify C. auris and differentiate it from C. haemulonii. It is inexpensive, fast and it could be a useful tool to reduce the chances of a C. auris outbreak. 相似文献2.
Jessie M. N. G. L. Suzuki Kenneth Osterhoudt Catiana H. Cartwright-Acar Destiny R. Gomez Sol Katzman Alan M. Zahler 《PLoS genetics》2022,18(2)
Pre-mRNA splicing is an essential step of eukaryotic gene expression carried out by a series of dynamic macromolecular protein/RNA complexes, known collectively and individually as the spliceosome. This series of spliceosomal complexes define, assemble on, and catalyze the removal of introns. Molecular model snapshots of intermediates in the process have been created from cryo-EM data, however, many aspects of the dynamic changes that occur in the spliceosome are not fully understood. Caenorhabditis elegans follow the GU-AG rule of splicing, with almost all introns beginning with 5’ GU and ending with 3’ AG. These splice sites are identified early in the splicing cycle, but as the cycle progresses and “custody” of the pre-mRNA splice sites is passed from factor to factor as the catalytic site is built, the mechanism by which splice site identity is maintained or re-established through these dynamic changes is unclear. We performed a genetic screen in C. elegans for factors that are capable of changing 5’ splice site choice. We report that KIN17 and PRCC are involved in splice site choice, the first functional splicing role proposed for either of these proteins. Previously identified suppressors of cryptic 5’ splicing promote distal cryptic GU splice sites, however, mutations in KIN17 and PRCC instead promote usage of an unusual proximal 5’ splice site which defines an intron beginning with UU, separated by 1nt from a GU donor. We performed high-throughput mRNA sequencing analysis and found that mutations in PRCC, and to a lesser extent KIN17, changed alternative 5’ splice site usage at native sites genome-wide, often promoting usage of nearby non-consensus sites. Our work has uncovered both fine and coarse mechanisms by which the spliceosome maintains splice site identity during the complex assembly process. 相似文献
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Epidemiology and Antifungal Susceptibilities of Yeasts Causing Vulvovaginitis in a Teaching Hospital
Soledad Gamarra Susana Morano Catiana Dudiuk Estefanía Mancilla María Elena Nardin Emilce de los Angeles Méndez Guillermo Garcia-Effron 《Mycopathologia》2014,178(3-4):251-258
Vulvovaginal candidiasis is one of the most common mycosis. However, the information about antifungal susceptibilities of the yeasts causing this infection is scant. We studied 121 yeasts isolated from 118 patients with vulvovaginal candidiasis. The isolates were identified by phenotypic and molecular methods, including four phenotypic methods described to differentiate Candida albicans from C. dubliniensis. Antifungal susceptibility testing was performed according to CLSI documents M27A3 and M27S4 using the drugs available as treatment option in the hospital. Diabetes, any antibacterial and amoxicillin treatment were statistically linked with vulvovaginal candidiasis, while oral contraceptives were not considered a risk factor. Previous azole-based over-the-counter antifungal treatment was statistically associated with non-C.albicans yeasts infections. The most common isolated yeast species was C. albicans (85.2 %) followed by C. glabrata (5 %), Saccharomyces cerevisiae (3.3 %), and C. dubliniensis (2.5 %). Fluconazole- and itraconazole-reduced susceptibility was observed in ten and in only one C. albicans strains, respectively. All the C. glabrata isolates showed low fluconazole MICs. Clotrimazole showed excellent potency against all but seven isolates (three C. glabrata, two S. cerevisiae, one C. albicans and one Picchia anomala). Any of the strains showed nystatin reduced susceptibility. On the other hand, terbinafine was the less potent drug. Antifungal resistance is still a rare phenomenon supporting the use of azole antifungals as empirical treatment of vulvovaginal candidiasis. 相似文献
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An autographic assay suitable for the detection of antioxidant compounds in a complex matrix (liquid and semi-solid pharmaceutical
formulations) or in isolated compounds was described. The pre-formed radical monocation of 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic
acid) (ABTS•+) was generated by oxidation of ABTS with potassium persulfate and reduced in the presence of hydrogen-donating antioxidants.
For a further comparative estimation of its applicability and sensitivity, different medicinal plant extracts, hydrogels and
antioxidant compounds were dot seeded or chromatographed on silica gel (TLC) and revealed with ABTS•+ solution (System I) or ABTS•+ immobilized by gel entrapment (System II). Both systems were effective and able to detect antioxidant activity in a micromolar
range in seconds. System II was more sensitive and reproducible than System I. This micromethod is quick, inexpensive, and
particularly helpful whether it works with numerous samples or on a small scale. 相似文献
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Catiana Dudiuk Laura Theill Soledad Gamarra Guillermo Garcia-Effron 《Current fungal infection reports》2017,11(4):176-183
Purpose of Review
The aim of this review is to evaluate these molecular-based methods able to identify pathogenic cryptic Candida spp. focusing on those that demonstrated to be useful in clinical laboratory settings.Recent Findings
It is long known that some Candida spp. are genetically heterogeneous. Firstly, individual species were divided into groups based on differences on the sequence of some genes. Later, those groups were designated as cryptic species and defined as phenotypically indistinguishable species that are only identified by their DNA sequences. Many common Candida spp. are now considered complexes formed by several cryptic species. Some of them have been recognized as human pathogens. The identification of these species is problematic but necessary since they have different host range, infection sites, infection severity, and antifungal susceptibility. Several independent DNA markers were proposed as tools for the differentiation of highly related species. We will concentrate on the three species complexes most frequently associated with human infections including Candida albicans, C. glabrata, and C. parapsilosis complexes and a fourth group of less common but multiresistant species including C. haeumulonii complex and C. auris.Summary
We review the clinically useful molecular tools able to differentiate the cryptic species of C. albicans, C. glabrata, and C. parapsilosis complexes and designated to uncover emerging multiresistant species.
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