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Registros recuperados: 8
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Adaptation of a real-time RT-PCR assay for the detection of Schmallenberg virus Rev Salud Anim.
Perera,Carmen L; Rios,Liliam; Frías,María T; Pérez,Lester J.
Schmallenberg virus was first detected in Germany in October 2011, associated with congenital malformations in cattle, sheep and goats. This novel emergent agent causes mild disease in cattle with decreased milk production, fever and diarrhea. In March 2012, the German Institute of Diagnostic Virology, Friedrich-Loeffler-Institut, reported the development and validation of a real-time RT-PCR for the diagnosis of this new virus. The Animal Virology Laboratory at the National Center for Animal and Plant Health in Cuba has adapted the protocol previously reported on the LightCycler platforms using two different mix conditions. In all cases, amplification curves obtained were specific and all the dilutions tested showed an increase in the Ct-values....
Tipo: Journal article Palavras-chave: Schmallenberg virus; Real-time RT-PCR; Emergent.
Ano: 2013 URL: http://scielo.sld.cu/scielo.php?script=sci_arttext&pid=S0253-570X2013000300009
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Cloning and expression analysis of rboh gene encoding respiratory burst oxidase in Citrullus colocynthis National Institute of Agronomic Research
Si, Y.; Kang, K.K.; Dane, F..
Drought stress is a major environmental factor influencing plant growth and development. Citrullus colocynthis is a very drought tolerant cucurbit species with a deep root system. Differences in gene expression during drought were studied using cDNA-AFLP. In this study, we focus on mRNA accumulation of drought-induced genes in C. colocynthis treated with PEG, abscisic acid (ABA), salicylic acid (SA), jasmonic acid (JA) or NaCl. Two genes, CcrbohD and CcrbohF, encoding respiratory burst oxidase proteins were cloned using RACE. Sequence comparison showed that the CcrbohD protein has a conserved region at the N terminus with an EF-hand loop domain, characteristic of the RBO protein family. Phylogenetic analysis confirmed that CcrbohD and CcrbohF belong to the...
Tipo: Poster Palavras-chave: Drought stress; PEG; Plant hormone; Real-time RT-PCR; Bitter apple; Citrullus colocynthis.
Ano: 2008 URL: http://hdl.handle.net/2174/269
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Evaluation of reference genes for real-time PCR studies of Brazilian Somalis sheep infected by gastrointestinal nematodes. Repositório Alice
ZAROS, L. G.; COUTINHO, L. L.; SIDER, L. H.; MEDEIROS, H. R. de; NEVES, M. R. M. das; BENVENUTI, C. L.; NAVARRO, A. M. do C.; VIEIRA, L. da S..
Abstract: Precise normalization with reference genes is necessary, in order to obtain reliable relative expression data in response to gastrointestinal nematode infection. By using sheep from temperate regions as models, three reference genes, viz., ribosomal protein LO (RPLO), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and succinate dehydrogenase complex subunit A (SDHA), were investigated in the abomasum, abomasal lymph nodes and small intestine of Brazilian Somalis sheep, either resistant or susceptible to gastrointestinal nematodes infections. Real time PCR was carried out by using SYBR Green I dye, and gene stability was tested by geNorm. RPLO was an ideal reference gene, since its expression was constant across treatments, presented lower...
Tipo: Artigo de periódico Palavras-chave: PCR em tempo real; Nematódeo gastrintestinal; Raça Somalis Brasileira; Control genes; Real-time RT-PCR; Gastrointestinal diseases; Nematodes.; Ovino; Parasitologia; Nematóide; Doença animal; Parasito de animal; Helminto gastrintestinal; Sheep; Nematode infections; Helminths; Parasites; Reverse transcriptase polymerase chain reaction..
Ano: 2010 URL: http://www.alice.cnptia.embrapa.br/alice/handle/doc/865496
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Evaluation of reference genes for real-time PCR studies of Brazilian Somalis sheep infected by gastrointestinal nematodes Genet. Mol. Biol.
Zaros,Lilian Giotto; Coutinho,Luiz Lehmann; Sider,Lúcia Helena; Medeiros,Henrique Rocha de; Neves,Maria Rosalba Moreira das; Benvenuti,Camila Loures; Navarro,Andrine Maria do Carmo; Vieira,Luiz da Silva.
Precise normalization with reference genes is necessary, in order to obtain reliable relative expression data in response to gastrointestinal nematode infection. By using sheep from temperate regions as models, three reference genes, viz., ribosomal protein LO (RPLO), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and succinate dehydrogenase complex subunit A (SDHA), were investigated in the abomasum, abomasal lymph nodes and small intestine of Brazilian Somalis sheep, either resistant or susceptible to gastrointestinal nematodes infections. Real time PCR was carried out by using SYBR Green I dye, and gene stability was tested by geNorm. RPLO was an ideal reference gene, since its expression was constant across treatments, presented lower variation, and...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Control genes; Gastrointestinal nematodes; Ovine; Real-time RT-PCR.
Ano: 2010 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572010000300018
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Evaluation of various real-time RT-PCR assays for the detection and quantitation of human norovirus ArchiMer
Butot, Sophie; Le Guyader, Soizick; Krol, Joanna; Putallaz, Thierry; Amoroso, Richard; Sanchez, Gloria.
Human noroviruses (NoVs) are the most common viruses causing acute gastroenteritis in humans. Performance characteristics of two commercial quantitative NoV RT-PCR assays, the Norovirus real-time RT-PCR Kit (AnDiaTec) and the Type I and Type II kits (Generon), and the international assay as selected by the CEN/TC/WG6/TAG4 group were evaluated for the specific detection and quantitation of 59 NoV samples, including different subtypes of NoV genogroup I and II. The results showed that the method proposed by the CEN/TC/WG6/TAG4 group was 100% specific since it was able to detect all samples tested. The commercialized kits evaluated failed to detect a vast majority of NoV GI strains. Additionally the Generon kit did not succeed to detect strains from GII.3,...
Tipo: Text Palavras-chave: Norovirus; Real-time RT-PCR; Commercial kit.
Ano: 2010 URL: http://archimer.ifremer.fr/doc/00006/11760/8473.pdf
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Limited replication of yellow fever 17DD and 17D-Dengue recombinant viruses in rhesus monkeys Anais da ABC (AABC)
Trindade,Gisela F.; Marchevsky,Renato S.; Fillipis,Ana M.B. de; Nogueira,Rita M.R.; Bonaldo,Myrna C.; Acero,Pedro C.; Caride,Elena; Freire,Marcos S.; Galler,Ricardo.
For the development of safe live attenuated flavivirus vaccines one of the main properties to be established is viral replication. We have used real-time reverse transcriptase-polymerase chain reaction and virus titration by plaque assay to determine the replication of yellow fever 17DD virus (YFV 17DD) and recombinant yellow fever 17D viruses expressing envelope proteins of dengue virus serotypes 2 and 4 (17D-DENV-2 and 17D-DENV-4). Serum samples from rhesus monkeys inoculated with YFV 17DD and 17D-DENV chimeras by intracerebral or subcutaneous route were used to determine and compare the viremia induced by these viruses. Viral load quantification in samples from monkeys inoculated by either route with YFV 17DD virus suggested a restricted capability of...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Yellow fever; Dengue; Vaccine; Attenuation; Replication; Real-time RT-PCR.
Ano: 2008 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652008000200009
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Quantification of bovine cytokine gene expression using real-time RT-PCR methodology Genet. Mol. Biol.
Zaros,Lilian Giotto; Bricarello,Patrízia Ana; Amarante,Alessandro Francisco Talamini; Coutinho,Luiz Lehmann.
T cells produce cytokines that affect host response to infection. This paper reports real-time RT-PCR conditions and validation steps for accurate quantification of Bos indicus cytokines, interleukin (IL)-2, IL-4, IL-8, IL12p-35, IL-13, tumoral necrosis factor (TNF)-alpha, interferon (IFN)-gamma, monocyte chemoattractant proteins (MCP)-1 and MCP-2, and the glycoprotein mucin (MUC)-1 in two groups of Nelore cattle, one resistant and the other susceptible to gastrointestinal nematode infections. RPL-19 was shown to be an ideal internal control gene, since its expression was constant across treatments and presented lower variation when compared to the GAPDH gene. The optimized conditions established in the present study can be used to determine the immune...
Tipo: Info:eu-repo/semantics/other Palavras-chave: Cytokine gene expression; Real-time RT-PCR; Cattle; Gastrointestinal nematodes; Bovine cytokines.
Ano: 2007 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572007000400012
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Validation of EN ISO method 15216-Part 1-Quantification of hepatitis A virus and norovirus in food matrices ArchiMer
Lowther, J. A.; Bosch, A.; Butot, S.; Ollivier, Joanna; Maede, D.; Rutjes, S. A.; Hardouin, G.; Lombard, B.; In'T Veld, P.; Leclercq, A..
Hepatitis A virus (HAV) and norovirus are important agents of food-borne human viral illness, with common vehicles including bivalve molluscan shellfish, soft fruit and various vegetables. Outbreaks of viral illness due to contamination of the surfaces of foods, or food preparation surfaces by for example infected food handlers are also common. Virus analysis of food matrices can contribute towards risk management for these hazards and a two-part technical specification for determination of Hepatitis A virus and norovirus in food matrices (ISO/TS 15216:2013) was published jointly by the European Committee for Standardisation and the International Organization for Standardization in 2013. As part of the European Mandate No. M381 to validate 15 standards in...
Tipo: Text Palavras-chave: Norovirus; Hepatitis A virus; Real-time RT-PCR; Validation; Standardization; Bivalve molluscan shellfish; Soft fruit; Vegetables; Bottled water; Surfaces.
Ano: 2019 URL: https://archimer.ifremer.fr/doc/00476/58776/61442.pdf
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