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Accuracy of quantitative polymerase chain reaction in samples of frozen and paraffin-embedded healthy skin for the diagnosis of canine visceral leishmaniasis Arq. Bras. Med. Vet. Zootec.
Campos,M.P.; Madeira,M.F.; Silva,D.A.; Solcà,M.S.; Espíndola,O.M.; Mendes Júnior,A.A.V.; Figueiredo,F.B..
ABSTRACT The purpose of the present work was to evaluate the accuracy of quantitative polymerase chain reaction (qPCR) performed on samples of fresh frozen tissue (FT) and formalin-fixed, paraffin-embedded (FFPE) healthy skin. This is a validation study conducted with samples from 46 dogs from an endemic area in Brazil. After sample collection, DNA extractions were conducted using commercial kits and qPCR was oriented to kinetoplast DNA (kDNA) targets of the Leishmania infantum species. The results obtained for the FFPE samples showed 63.6% sensitivity and 77.1% specificity, whereas those obtained for the FT samples showed 100% and 48.6%, respectively. Poor agreement was observed for the results of the qPCR technique with FT and FFPE samples. Our results...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Diagnosis; QPCR; Canine visceral leishmaniasis.
Ano: 2017 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352017000601443
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An updated assessment of Symbiodinium spp. that associate with common scleractinian corals from Moorea (French Polynesia) reveals high diversity among background symbionts and a novel finding of clade B ArchiMer
Rouze, Heloise; Lecellier, Gael J.; Saulnier, Denis; Planes, Serge; Gueguen, Yannick; Wirshing, Herman H.; Berteaux-lecellier, Veronique.
The adaptative bleaching hypothesis (ABH) states that, depending on the symbiotic flexibility of coral hosts (i.e., the ability of corals to “switch” or “shuffle” their algal symbionts), coral bleaching can lead to a change in the composition of their associated Symbiodinium community and, thus, contribute to the coral’s overall survival. In order to determine the flexibility of corals, molecular tools are required to provide accurate species delineations and to detect low levels of coral-associated Symbiodinium. Here, we used highly sensitive quantitative (real-time) PCR (qPCR) technology to analyse five common coral species from Moorea (French Polynesia), previously screened using only traditional molecular methods, to assess the presence of...
Tipo: Text Palavras-chave: Corals; French polynesia; Clade B; Symbiodinium; QPCR; Flexibility; Generalist; Faithful clade.
Ano: 2017 URL: http://archimer.ifremer.fr/doc/00365/47652/47683.pdf
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Associação da carga parasitária renal com achados laboratoriais em cães com leishmaniose visceral Ciência Rural
Torres,Mariana de Medeiros; Almeida,Arleana do Bom Parto Ferreira de; Sorte,Eveline da Cruz Boa; Paula,Daphine Ariadne Jesus de; Oliveira,Anderson Castro Soares de; Pescador,Caroline Argenta; Mendonça,Adriane Jorge; Nakazato,Luciano; Sousa,Valéria Régia Franco.
A leishmaniose visceral canina é uma doença grave e a morte ocorre por falência renal, considerando que os métodos diagnósticos convencionais não possibilitam a classificação clínica do animal. O objetivo deste estudo foi associar a carga parasitária renal aos achados clínicos e histopatológicos em cães com leishmaniose visceral. A análise microscópica revelou predomínio de nefrite intersticial mononuclear de graus variados em 59,3% dos cães avaliados. Entretanto, não houve diferença entre a carga parasitária renal de sintomáticos e oligossintomáticos (P= 0,35). As lesões renais foram de ordem inflamatória e a quantidade de parasitos não influenciaram na característica dessas lesões e nem nas alterações bioquímicas, mesmo em cães com diferentes...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Rim; QPCR; Cão; Leishmania sp..
Ano: 2013 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782013000500023
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Avaliação da degradação do DNA por PCR em tempo real em embriões de soja envelhecida artificialmente. Repositório Alice
LOPES, R. M.; CARVALHO, P. A. S. de V.; BRASILEIRO, A. C. M.; SARAIVA, M. A. de P.; GIMENES, M. A..
2016
Tipo: Resumo em anais de congresso (ALICE) Palavras-chave: Dano DNA; QPCR; Semente.
Ano: 2016 URL: http://www.alice.cnptia.embrapa.br/handle/doc/1067931
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Babesia bovis and Babesia bigemina infection levels estimated by qPCR in Angus cattle from an endemic area of São Paulo state, Brazil. Repositório Alice
GIGLIOTI, R.; OLIVEIRA, H. N. de; SANTANA, C. H.; IBELLI, A. M. G.; NEO, T. A.; BILHASSI, T. B.; RABELO, M. D.; MACHADO, R. Z.; BRITO, L. G.; OLIVEIRA, M. C. de S..
The levels of infection by Babesia bovis and Babesia bigemina were estimated by absolute quantification through the quantitative PCR technique (qPCR). Fifty-one contemporaneous Angus cattle were evaluated on two occasions. The number of standard female Rhipicephalus microplus ticks present on the left side of the body was counted and blood samples were drawn from the tail vein into tubes containing the anticoagulant EDTA. The blood samples were submitted to DNA extraction and used to quantify the number of copies (NC) of DNA from B. bovis and B. bigemina by qPCR. The data on tick count and number of DNA copies were transformed for normalization and analyzed by a mixed model method. A multivariate model with repeated measures of the same animal, including...
Tipo: Artigo em periódico indexado (ALICE) Palavras-chave: Babesiosis; R microplus; Resistance; QPCR; Bovino; Babesia bovis.
Ano: 2016 URL: http://www.alice.cnptia.embrapa.br/handle/doc/1057560
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Biodiversity and quantification of functional bacteria in deammonification process. Repositório Alice
PRÁ, M. C. de; KUNZ, A.; BORTOLI, M.; VIANCELLI, A.; SCUSSIATO, L. A.; SOARES, H. M..
2015
Tipo: Artigo em anais de congresso (ALICE) Palavras-chave: ANAMMOX; Ammonia oxidizing bacteria; Partial nitritation; FISH; QPCR.
Ano: 2015 URL: http://www.alice.cnptia.embrapa.br/handle/doc/1045554
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Chilean IPNV isolates: Robustness analysis of PCR detection Electron. J. Biotechnol.
Jorquera,Esteban; Morales,Paz; Tapia,David; Torres,Pamela; Eissler,Yoanna; Espinoza,Juan C; Conejeros,Pablo; Kuznar,Juan.
Background: The genomes of several infectious pancreatic necrosis viruses (IPNVs) isolated in Chile were sequenced with a single amplification approach for both segments A and B. The resulting sequences were then used to determine the conservation of the primer-binding regions used in polymerase chain reaction (PCR)-based diagnostic methods proposed in the literature. Thus, the robustness of each technique was studied, particularly the eventual effect of further mutations within the primer-binding sites. Results: On analysis, most methods currently used to detect Chilean IPNV varieties were deemed adequate. However, the primers were designed to be genogroup specific, implying that most detection methods pose some risk of detecting all strains prevalent in...
Tipo: Journal article Palavras-chave: IPNV detection; Mismatch's Tm analysis; QPCR.
Ano: 2016 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000200005
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Ciguatoxin Occurrence in Food-Web Components of a Cuban Coral Reef Ecosystem: Risk-Assessment Implications ArchiMer
Díaz-asencio, Lisbet; Clausing, Rachel J.; Vandersea, Mark; Chamero-lago, Donaida; Gómez-batista, Miguel; Hernández-albernas, Joan I.; Chomérat, Nicolas; Rojas-abrahantes, Gabriel; Litaker, Wayne; Tester, Patricia; Diogène, Jorge; Alonso-hernández, Carlos M.; Dechraoui Bottein, Marie-yasmine.
In Cuba, ciguatera poisoning associated with fish consumption is the most commonly occurring non-bacterial seafood-borne illness. Risk management through fish market regulation has existed in Cuba for decades and consists of bans on selected species above a certain weight; however, the actual occurrence of ciguatoxins (CTXs) in seafood has never been verified. From this food safety risk management perspective, a study site locally known to be at risk for ciguatera was selected. Analysis of the epiphytic dinoflagellate community identified the microalga Gambierdiscus. Gambierdiscus species included six of the seven species known to be present in Cuba (G. caribaeus, G. belizeanus, G. carpenteri, G. carolinianus, G. silvae, and F. ruetzleri). CTX-like...
Tipo: Text Palavras-chave: Caribbean; Ciguatoxicity; QPCR; Trophic transfer; Ish; Food safety; Food security; Science-based management; Foodborne disease.
Ano: 2019 URL: https://archimer.ifremer.fr/doc/00597/70947/69189.pdf
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Clonagem e análise de expressão de candidatos a genes de referência em tambaqui (Colossoma macropomum). Repositório Alice
NASCIMENTO, A. R. do; O'SULLIVAN, F. A.; SILVA, G. F. da.
Neste trabalho, os genes 18S, beta-actina, gapdh e ef1alfa do tambaqui foram clonados e sequenciados, visando à obtenção e validação de genes de referência, candidatos aos estudos genômicos e de fisiologia na espécie.
Tipo: Resumo em anais de congresso (ALICE) Palavras-chave: QPCR; Normalizador; Expressão gênica; Normfinder.
Ano: 2016 URL: http://www.alice.cnptia.embrapa.br/handle/doc/1047706
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Comparative validation using quantitative real-time PCR (qPCR) and conventional PCR of bovine semen centrifuged in continuous density gradient Arq. Bras. Med. Vet. Zootec.
Resende,M.V.; Lucio,A.C.; Perini,A.P.; Oliveira,L.Z.; Almeida,A.O.; Alves,B.C.A.; Moreira-Filho,C.A.; Santos,I.W.; Hossepian de Lima,V.F.M..
The objective of the present study was to determine the sperm enrichment with X-bearing spermatozoa, after one centrifugation in a Percoll or OptiPrep continuous density gradient, using quantitative real-time polymerase chain reaction (qPCR) of sperm DNA and resultant in vitro-produced bovine embryos by PCR. Frozen/thawed sperm was layered on density gradients and the tubes were centrifuged. Supernatants were gently aspirated and the sperm recovered from the bottom of the tubes. Cleavage and blastocyst rates were determined through in vitro production of embryos and PCR was performed to identify the embryos' genetic sex. A difference in blastocyst rate was found in the Percoll treatment compared to OptiPrep (P<0.05). The percentage of female embryos in...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Bovine; Sperm sexing; Centrifugation; Embryo sexing; QPCR; PCR.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352011000300002
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Correlation between chronic inflammation, immunostaining and parasite load in the genital system of female dogs naturally infected with Leishmania infantum Ciência Rural
Oliveira,Vinícius Vasconcelos Gomes de; Macedo,Simone Regina Barros de; Araújo,Flábio Ribeiro de; Ramos,Carlos Alberto do Nascimento; Ramos,Rafael Antonio Nascimento; Maia,Frederico Celso Lyra; Alves,Leucio Câmara; Silva Junior,Valdemiro Amaro da.
ABSTRACT: The aim of the present study was to investigate the correlations among chronic inflammatory reaction, immunostaining and parasite load in the genital system of female dogs naturally infected with Leishmania infantum . Animals (n = 10) used in this study were from the Department of Vector Control and Animal Surveillance of the municipality of Caruaru, state of Pernambuco, Brazil. Fragments of the vulva, vagina, cervix, uterine body, uterine horns and ovaries were submitted to histopathological analysis, immunohistochemistry (IHC) and DNA detection of amastigotes by qPCR. Correlations were found between the IHC findings and chronic inflammatory infiltrate related to L. Infantum only in the vulva and vagina; whereas, the same inflammatory reactions...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Canine visceral leishmaniasis; Genital pathology; Immunohistochemistry; QPCR.
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782016001102029
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Cuantificación de maíz genéticamente modificado mediante las técnicas de qPCR y dPCR Agrociencia
Gutiérrez-Angoa,Lizbeth E.; Castillo-Durán,Luis C.; Gómez-Castelo,Blanca E.; Acatzi-Silva,Abraham I..
A partir de la emisión de los permisos de liberación al ambiente de maíz (Zea mays L.) genéticamente modificado (GM) en el año 2009, en México fue necesario realizar la detección, identificación y cuantificación de organismos genéticamente modificados (OGM) de los cultivos en el país. El objetivo del presente estudio fue validar la técnica de cuantificación absoluta de mezclas de hoja en fracción masa de maíz GM a través de la técnica de la reacción en cadena de la polimerasa (PCR) en tiempo real (qPCR) y digital (dPCR). Mezclas de hojas de maíz modificado con el evento MON810 se prepararon con hojas de maíz convencional, fueron analizadas con la técnica de qPCR, y como calibrantes se usaron un plásmido de referencia certificado y ADN obtenido de una hoja...
Tipo: Info:eu-repo/semantics/article Palavras-chave: QPCR; DPCR; Zea mays L.; OGM.
Ano: 2015 URL: http://www.scielo.org.mx/scielo.php?script=sci_arttext&pid=S1405-31952015000400003
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Detecção de Babesia bovis por qPCR em bezerros leiteiros recém-nascidos. Repositório Alice
GONÇALVES, T. C.; FANTATTO, R. R.; FEITOSA, L. A.; DOMINGUES, L. F.; NÉO, T. A.; GIGLIOTI, R.; BILHASSI, T.; RABELO, M. D.; SANTANA, R. C. M.; CHAGAS, A. C. de S.; OLIVEIRA, M. C. de S..
2012
Tipo: Resumo em anais de congresso (ALICE) Palavras-chave: Babasea bovis; QPCR; Detecção.
Ano: 2012 URL: http://www.alice.cnptia.embrapa.br/handle/doc/946146
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Detecção de células viáveis de patógenos em queijo Minas Padrão pela técnica de PCR em Tempo Real. Repositório Alice
MENDONÇA, J. F. M. de; VIEIRA, F. de O.; FONSECA, I.; ARCURI, E. F.; RIBEIRO, J. B.; BORGES, M. de F.; MARTINS, M. F..
2015
Tipo: Resumo em anais de congresso (ALICE) Palavras-chave: EMA; Micro-organismos patogênicos; Derivados lácteos; QPCR; EMA; Micro-organismos patogênicos; Derivados lácteos; QPCR; Viabilidade.
Ano: 2015 URL: http://www.alice.cnptia.embrapa.br/handle/doc/1029283
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Detection and quantification of human adenovirus genomes in Acanthamoeba isolated from swimming pools Anais da ABC (AABC)
STAGGEMEIER,RODRIGO; ARANTES,THALITA; CAUMO,KARIN S.; ROTT,MARILISE B.; SPILKI,FERNANDO R..
ABSTRACT Acanthamoeba is the most common free-living environmental amoeba, it may serve as an important vehicle for various microorganisms living in the same environment, such as viruses, being pathogenic to humans. This study aimed to detect and quantify human adenoviruses (HAdV) in Acanthamoebas isolated from water samples collected from swimming pools in the city of Porto Alegre, Southern Brazil. Free-living amoebae of the genus Acanthamoeba were isolated from water samples, and isolates (n=16) were used to investigate the occurrence of HAdVs. HAdV detection was performed by quantitative real-time polymerase chain reaction (qPCR). HAdVs were detected in 62.5% (10/16) of Acanthamoeba isolates, ranging from 3.24x103 to 5.14x105 DNA copies per milliliter...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Acanthamoeba; Free-living amoebae; Human adenovirus; QPCR; Swimming pools.
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652016000200635
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Detection of Babesia bovis and Babesia bigemina in water buffaloes (Bubalus bubalis) in endemic areas of São Paulo state, Brazil. Repositório Alice
NEO, T. A.; GIGLIOTI, R.; OBREGON, D.; BILHASSI, T. B.; OLIVEIRA, H. N. de; MACHADO, R. Z.; ANIBAL, F. de F.; BRITO, L. G.; MALAGO JUNIOR, W.; DONATONI, F. A. B.; OLIVEIRA, M. C. de S..
bitstream/item/150906/1/OJVM-2016052415273391.pdf
Tipo: Artigo em periódico indexado (ALICE) Palavras-chave: Diagnosis; NPCR; QPCR; Bufalo; Bubalus Bubalis; Buffaloes; Babesiosis.
Ano: 2016 URL: http://www.alice.cnptia.embrapa.br/alice/handle/doc/1057566
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Detection of Salmonella spp. by Conventional Bacteriology and by Quantitative Polymerase-Chain Reaction in Commercial Egg Structures Rev. Bras. Ciênc. Avic.
Moraes,DMC; Duarte,SC; Bastos,TSA; Rezende,CLG; Leandro,NSM; Café,MB; Stringhini,JH; Andrade,MA.
Abstract Conventional bacteriology techniques and quantitative polymerase-chain reaction (qPCR) were applied to the eggshell, albumen, and yolk of washed and unwashed commercial white and brown eggs, to detect Salmonella spp. Pooled samples of eggshells, albumen, and yolk of white and brown eggs were collected at the poultry house and at the egg-storage room. Salmonella spp. was detected by conventional bacteriology in 5.4% (21/387) of analyzed samples and in 16% (68/387) by qPCR. In the 114 unwashed white eggs samples of eggshell, albumen and yolk, the bacterium was identified in 2.6% of the eggs (3/114) by conventional bacteriology and in 13.2% (15/114) by qPCR. In the 90 samples of washed eggs, 6.7% (6/90) were contaminated as detected by conventional...
Tipo: Info:eu-repo/semantics/article Palavras-chave: QPCR; Eggshells; Albumen; Yolk.
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-635X2016000100117
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Development and validation of a modified TaqMan based real-time PCR assay targeting the lipl32 gene for detection of pathogenic Leptospira in canine urine samples BJM
Miotto,Bruno Alonso; Hora,Aline Santana da; Taniwaki,Sueli Akemi; Brandão,Paulo Eduardo; Heinemann,Marcos Bryan; Hagiwara,Mitika Kuribayashi.
Abstract A modified TaqMan real-time polymerase chain reaction targeting a 138 bp fragment within the lipl32 gene was developed to identify exclusively pathogenic Leptospira spp. in dog urine samples. Thirty-five samples from dogs with suspected clinical leptospirosis and 116 samples from apparently healthy dogs were tested for presence of leptospiral DNA using the TaqMan-based assay. The results were compared with those from a well-established conventional PCR targeting the 16S RNA encoding gene associated with nucleotide sequencing analysis. The overall agreement between the assays was 94.8% (confidence interval [CI] 95% 88-100%). The newly developed assay presented 91.6% (CI 95% 71.5-98.5%) relative sensitivity (22[+] lipl32 PCR/24[+] 16S RNA and...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Leptospirosis; Dog; Urine; QPCR; Lipl32.
Ano: 2018 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822018000300584
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Development of a qPCR Method for the Identification and Quantification of Two Closely Related Tuna Species, Bigeye Tuna ( Thunnus obesus ) and Yellowfin Tuna ( Thunnus albacares ), in Canned Tuna ArchiMer
Bojolly, Daline; Doyen, Perine; Le Fur, Bruno; Christaki, Urania; Verrez-bagnis, Veronique; Grard, Thierry.
Bigeye tuna (Thunnus obesus) and yellowfin tuna (Thunnus albacares) are among the most widely used tuna species for canning purposes. Not only substitution but also mixing of tuna species is prohibited by the European regulation for canned tuna products. However, as juveniles of bigeye and yellowfin tunas are very difficult to distinguish, unintentional substitutions may occur during the canning process. In this study, two mitochondrial markers from NADH dehydrogenase subunit 2 and cytochrome c oxidase subunit II genes were used to identify bigeye tuna and yellowfin tuna, respectively, utilizing TaqMan qPCR methodology. Two different qPCR-based methods were developed to quantify the percentage of flesh of each species used for can processing. The first one...
Tipo: Text Palavras-chave: Tuna; Authentication; Quantification; Canned products; TaqMan; QPCR; Species identification; Bigeye tuna (Thunnus obesus); Yellowfin tuna (Thunnus albacares).
Ano: 2017 URL: https://archimer.ifremer.fr/doc/00371/48217/48403.pdf
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Development of methodologies for virus detection in soybean and wheat seeds. Repositório Alice
BOTELHO, S. R. A.; MARTINS, T. P.; DUARTE, M. F.; BARBOSA, A. V.; LAU, D.; FERNANDES, F. R.; SANCHES, M. M..
2016
Tipo: Artigo em periódico indexado (ALICE) Palavras-chave: SMV; WSMV; Quarantine; Seed certification; RT-PCR; QPCR; Quarentena; Certificação de semente; Virose; Trigo; Soja.
Ano: 2016 URL: http://www.alice.cnptia.embrapa.br/handle/doc/1036027
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