PCR-AGE, automated and manual methods to identify Candida strains from veterinary sources: A comparative approach

Brito, Érika Helena Salles de; Brilhante, Raimunda Sâmia Nogueira; Cordeiro, Rossana de Aguiar; Sidrim, J. J. C.; Fontenelle, Raquel Oliveira dos Santos; Melo, Luciana Magalhães; Albuquerque, Erica S.; Rocha, Marcos F.G.

doi:10.1016/j.vetmic.2009.06.031

Cited by 14 publications

(18 citation statements)

References 15 publications

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“…According to Chen et al, 54 the ITS1 and ITS2 regions are useful to differentiate closely related species, particularly those with identical D1/D2 domain DNA sequences. In addition, these regions distinguish yeast strains exhibiting differences of 10 bp or more in terms of PCR product lengths 55 . Thus, species identification accuracy should depend on both the D1/D2 domain of the large subunit of 26S rDNA as well as the ITS1 and ITS2 regions.…”

Section: Discussionmentioning

confidence: 99%

High-temperature ethanol production using thermotolerant yeast newly isolated from Greater Mekong Subregion

Techaparin

Thanonkeo

Klanrit

2017

Brazilian Journal of Microbiology

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The application of high-potential thermotolerant yeasts is a key factor for successful ethanol production at high temperatures. Two hundred and thirty-four yeast isolates from Greater Mekong Subregion (GMS) countries, i.e., Thailand, The Lao People's Democratic Republic (Lao PDR) and Vietnam were obtained. Five thermotolerant yeasts, designated Saccharomyces cerevisiae KKU-VN8, KKU-VN20, and KKU-VN27, Pichia kudriavzevii KKU-TH33 and P. kudriavzevii KKU-TH43, demonstrated high temperature and ethanol tolerance levels up to 45 °C and 13% (v/v), respectively. All five strains produced higher ethanol concentrations and exhibited greater productivities and yields than the industrial strain S. cerevisiae TISTR5606 during high-temperature fermentation at 40 °C and 43 °C. S. cerevisiae KKU-VN8 demonstrated the best performance for ethanol production from glucose at 37 °C with an ethanol concentration of 72.69 g/L, a productivity of 1.59 g/L/h and a theoretical ethanol yield of 86.27%. The optimal conditions for ethanol production of S. cerevisiae KKU-VN8 from sweet sorghum juice (SSJ) at 40 °C were achieved using the Box–Behnken experimental design (BBD). The maximal ethanol concentration obtained during fermentation was 89.32 g/L, with a productivity of 2.48 g/L/h and a theoretical ethanol yield of 96.32%. Thus, the newly isolated thermotolerant S. cerevisiae KKU-VN8 exhibits a great potential for commercial-scale ethanol production in the future.

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Section: Discussionmentioning

confidence: 99%

High-temperature ethanol production using thermotolerant yeast newly isolated from Greater Mekong Subregion

Techaparin

Thanonkeo

Klanrit

2017

Brazilian Journal of Microbiology

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“…In contrast, veterinary yeast isolates can be cultured from a wide variety of animal species, allowing for the possibility of more diversity among the isolates identified. Though much work has been performed examining phenotypic and genotypic methods of yeast identification from veterinary sources, the majority of these studies have concentrated on a single species of either animal or yeast (2,15,16,28,29). To our knowledge, there has been only one large-scale examination of multiple yeast and animal species to assess yeast identification (5).…”

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confidence: 99%

Molecular Identification of Veterinary Yeast Isolates by Use of Sequence-Based Analysis of the D1/D2 Region of the Large Ribosomal Subunit

et al. 2010

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Conventional methods of yeast identification are often time-consuming and difficult; however, recent studies of sequence-based identification methods have shown promise. Additionally, little is known about the diversity of yeasts identified from various animal species in veterinary diagnostic laboratories. Therefore, in this study, we examined three methods of identification by using 109 yeast samples isolated during a 1-year period from veterinary clinical samples. Comparison of the three methods-traditional substrate assimilation, fatty acid profile analysis, and sequence-based analysis of the region spanning the D1 and D2 regions (D1/D2) of the large ribosomal subunit-showed that sequence analysis provided the highest percent identification among the three. Sequence analysis identified 87% of isolates to the species level, whereas substrate assimilation and fatty acid profile analysis identified only 54% and 47%, respectively. Less-stringent criteria for identification increased the percentage of isolates identified to 98% for sequence analysis, 62% for substrate assimilation, and 55% for fatty acid profile analysis. We also found that sequence analysis of the internal transcribed spacer 2 (ITS2) region provided further identification for 36% of yeast not identified to the species level by D1/D2 sequence analysis. Additionally, we identified a large variety of yeast from animal sources, with at least 30 different species among the isolates tested, and with the majority not belonging to the common Candida spp., such as C. albicans, C. glabrata, C. tropicalis, and the C. parapsilosis group. Thus, we determined that sequence analysis of the D1/D2 region was the best method for identification of the variety of yeasts found in a veterinary population.In both veterinary and human diagnostic laboratories, correct identification of yeasts is important for the care of patients. Traditionally, yeast identification has been performed using biochemical analysis, substrate assimilation methods, morphological examination, or various combinations of the three. To increase the ease of identification, commercial tests that use these methods have been created, but despite the convenience provided by these methods, identification of yeasts by these conventional applications can still be timeconsuming and difficult. In addition, considerable variability in the efficacy of these methods has been reported for identification of clinically important yeast (11; also reviewed in references 12, 32, 36, and 42), attributed primarily to the limitations of the databases used for the comparison of clinical isolates, as well as the subjectivity involved in the interpretation of results. Recent studies have also examined the effectiveness of various molecular identification methods for yeasts by the use of rRNA genes, with the internal transcribed spacer 1 (ITS1) and ITS2 regions and the region spanning the D1 and D2 regions (D1/ D2) shown to be the most useful for species-level identification of yeasts, as a result of the variability within...

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“…Fisher's exact test was applied to analyze the correspondence (P < 0.05) among manual identification method, API 20C, VITEK 2 and PCR-REA. The manual method was considered the gold standard (Brito et al, 2009).…”

Section: Discussionmentioning

confidence: 99%

PCR-REA as an important tool for the identification of Cryptococcus neoformans and Cryptococcus gattii from human and veterinary sources

Cordeiro

Costa

Brilhante

et al. 2011

Veterinary Microbiology

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The extraordinary ability of Cryptococcus species to cause disease has focused the attention of scientists on finding ways to improve their identification methods. In this study, PCR-REA, manual methods (morphological and biochemical characteristics), API 20C and VITEK 2 were used to test identify a total of 30 Cryptococcus spp. from human and veterinary sources. PCR-REA was performed using the capsular region as amplification target followed by restriction with the enzymes AgeI, BsmFI and HpaII. PCR-REA identified the strains as C. neoformans var. grubii (n=19) and C. gattii (n=8). There was no significant difference between the API 20C AUX and VITEK 2 when compared to manual methods for the identification of Cryptococcus spp. However, none of these non-manual methods were able to detect C. gattii samples. PCR-REA showed a greater level of concordance with the manual method, besides being faster and more sensitive than the other methods. Therefore, it is indicated for routine identification of Cryptococcus spp. strains.

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PCR-AGE, automated and manual methods to identify Candida strains from veterinary sources: A comparative approach

Cited by 14 publications

References 15 publications

High-temperature ethanol production using thermotolerant yeast newly isolated from Greater Mekong Subregion

High-temperature ethanol production using thermotolerant yeast newly isolated from Greater Mekong Subregion

Molecular Identification of Veterinary Yeast Isolates by Use of Sequence-Based Analysis of the D1/D2 Region of the Large Ribosomal Subunit

PCR-REA as an important tool for the identification of Cryptococcus neoformans and Cryptococcus gattii from human and veterinary sources

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