Rapid methods to extract DNA and RNA from<i>Cryptococcus neoformans</i>

Bolano, Alessandro; Stinchi, Silvia; Preziosi, Roberta; Bistoni, Francesco; Allegrucci, M.; Baldelli, Franco; Martini, Alessandro; Cardinali, Gianluigi

doi:10.1111/j.1567-1364.2001.tb00037.x

Cited by 60 publications

(32 citation statements)

References 12 publications

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“…Extração do DNA Genômico: A extração de DNA genômico foi realizada conforme descrito por BOLANO et al (2001) com algumas adaptações. O protocolo baseia-se na lise mecânica das células, obtida pelo uso de pérolas de vidro tratadas e foi adaptado para o gênero Staphylococcus, não sendo necessária a adição de enzimas para a ruptura da parede celular e membrana plasmática.…”

Section: Methodsunclassified

PERFIL DE RESISTÊNCIA A ANTIMICROBIANOS DE Staphylococcus aureus ISOLADOS DE GRANJAS E FRIGORÍFICOS DE SUÍNOS

Masson

Ferreira

Carvalho

2012

AVS

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RESUMO:Microrganismos da espécie S. aureus são responsabilizados por diversos problemas clínicos em suinocultura e humanos. Estudos epidemiológicos comprovam o potencial deste microrganismo em adquirir resistência a antibióticos. Atualmente, estirpes resistentes a meticilina conhecida pela sigla MRSA derivada do inglês (Meticilin Resistant Staphylococcus aureus), responsabilizados por casos de infecções nosocomiais, são as mais estudadas uma vez que o MRSA encontra-se disseminado em ambientes extra-hospitalares e frenquentemente tem sido isolado de vários animais domésticos inclusive suínos. O objetivo desde trabalho foi determinar a presença de S. aureus em granjas de suínos, identificar a ocorrência dos genes mecA, icaA e icaD e o perfil de resistência a antimicrobianos. Ao todo colheram-se 458 amostras de cinco granjas e dois frigoríficos. As amostras foram semeadas em ágar Braid -Parker e ágar sangue seguido de provas bioquímicas. As amostras sugestivas, foram submetidas a PCR para confirmação de espécie, detecção do gene coa para confirmação do gene da coagulase, mecA para avaliar a resistência a meticilina além dos genes de virulência icaA e icaD que expressam capacidade para formação de biofilmes. Na sequência, realizou-se o antibiograma para a avaliação de 11 antimicrobianos. Ao todo foram identificados 81 (79%) S. aureus isolados de todas as granjas e frigoríficos incluindo quatro amostras isoladas de funcionários das granjas. Nenhuma amostra foi positiva para o gene mecA. Em relação aos genes icaA e icaD, observou predomínio do gene icaD e que 41% das amostras foram positivas para os dois genes. O antibiograma demonstrou grande resistência às penicilinas e tetraciclinas, além de grande quantidade de S aureus multirresistentes. Palavras-chave: antibiograma; biofilmes; MRSA; suínos; staphylococcus; resistência ANTIBIOTIC RESISTANT PROFILE OF STAPHYLOCOCCUS AUREUS ISOLATED FROM PIGS FARMS AND SLAUTHERHOUSESABSTRACT: Staphylococcus aureus are involved in a wide range of clinical problems to swine industry as son in humans. Epidemiological researchs prove his potential to acquire resistantence to antibiotics. Nowadays, methicillin-resistant S. aureus (MRSA) are responsabilized for nosocomial infections and many studies are done because MRSA are spread to extra hospitalar enrivonment and frequentely isolated from domestic animals including pigs. The aim of this study was to determine the presence o S. aureus at swine farms and identify the mecA, icaA and icaD genes and the resistant proflife to antibiotics. Overal, 458 swabs were taked from five pigeris and two slautherhouses. All the samples were placed on Braid -Parker and blood agar follow by biochemical analyses. The suspect colonies were submitted to PCR to confirm the S. aureus species, by the detection of the coa gene, mecA to avaible meticillin-resistant as son to the virulence gens icaA and icaD that can determine slime production. Antibiogram were done to evaluate the response to 11 antibiotics. All pigeris and slautherhouse were positive and 81 (7...

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Section: Methodsunclassified

PERFIL DE RESISTÊNCIA A ANTIMICROBIANOS DE Staphylococcus aureus ISOLADOS DE GRANJAS E FRIGORÍFICOS DE SUÍNOS

Masson

Ferreira

Carvalho

2012

AVS

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“…DNA was extracted as previously described (Bolano et al 2001;Cardinali et al 2002) and amplified on a Hybaid Statistical analyses of electrophoretic data first requires that identical bands be located in different lanes and on separate gels. This was done using existing methodology (Cardinali et al 2002), which can be briefly outlined as follows.…”

Section: Rapdmentioning

confidence: 99%

Use of RAPD and killer toxin sensitivity in Saccharomyces cerevisiae strain typing

et al. 2005

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Aims: Two different strain characterization techniques, random amplified polymorphic DNA (RAPD) and killer toxin sensitivity (KTS), were compared to assess their typing performance using a set of 30 certified Saccharomyces cerevisiae strains. Methods and Results: A sequential random resampling procedure was employed to subdivide the 32 descriptors in eight sets, in order to compare the differential performances of the two techniques with diverse number of characters. Results showed that RAPD performs better than killer, although the complete differentiation of the strains under study could be obtained only by combining profiles from the two techniques Conclusions: The combination of different typing techniques was useful when discriminating similar organisms. In such cases, the introduction of a second typing technique can be more advantageous than increasing the number of characters obtained with a single method. Significance and Impact of the Study: The distribution of among-strains pairwise distances and the relative performance of the two techniques has implications for the study of biodiversity, taxonomy and microbial ecology.

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“…Twenty-two out of 35 strains were ascomycetous yeasts and 13 were basidiomycetous yeasts. The identifications of all strains were verified by sequence analyses of internal transcribed spacers (ITS) 1 and 2 and the D1/D2 regions of large subunit ribosomal DNA (rDNA) (see Table S1) (12,25). All strains were different from those present in the commercial (BDAL) database, but in an extended (BDALϩCBS in-house) database reference, the MSP of seven out of the 35 send strains were present (see Table S1).…”

Section: Strains and Sample Preparation For Maldi-tof Ms Analysismentioning

confidence: 99%

Interlaboratory Comparison of Sample Preparation Methods, Database Expansions, and Cutoff Values for Identification of Yeasts by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Using a Yeast Test Panel

et al. 2014

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An interlaboratory study using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) to determine the identification of clinically important yeasts (n ‫؍‬ 35) was performed at 11 clinical centers, one company, and one reference center using the Bruker Daltonics MALDI Biotyper system. The optimal cutoff for the MALDI-TOF MS score was investigated using receiver operating characteristic (ROC) curve analyses. The percentages of correct identifications were compared for different sample preparation methods and different databases.

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Rapid methods to extract DNA and RNA fromCryptococcus neoformans

Cited by 60 publications

References 12 publications

PERFIL DE RESISTÊNCIA A ANTIMICROBIANOS DE Staphylococcus aureus ISOLADOS DE GRANJAS E FRIGORÍFICOS DE SUÍNOS

PERFIL DE RESISTÊNCIA A ANTIMICROBIANOS DE Staphylococcus aureus ISOLADOS DE GRANJAS E FRIGORÍFICOS DE SUÍNOS

Use of RAPD and killer toxin sensitivity in Saccharomyces cerevisiae strain typing

Interlaboratory Comparison of Sample Preparation Methods, Database Expansions, and Cutoff Values for Identification of Yeasts by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Using a Yeast Test Panel

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