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Canine ehrlichiosis: clinical, hematological, serological and molecular aspects Ciência Rural
Nakaghi,Andréa Cristina Higa; Machado,Rosangela Zacarias; Costa,Mirela Tinucci; André,Marcos Rogério; Baldani,Cristiane Divan.
The aim of the present study was to compare the direct detection methods of Ehrlichia canis (blood smears and nested PCR), serological tests (Dot-ELISA and Immunofluorescent Antibody Test - IFAT), and demonstrate the most suitable test for the diagnosis of different stages of infection. Blood samples and clinical data were collected from 30 dogs examined at the Veterinary Teaching Hospital, UNESP, Jaboticabal, SP, Brazil. The clinical signs most frequently observed were apathy, anorexia, pale mucous membrane, fever, lymphadenopathy, splenomegaly, hemorrhages and uveitis. Evaluating the humoral immune response, 63.3% of the sera were IFAT positive, while 70% were Dot-ELISA positive. By nestedPCR 53.3% of the samples were positive. Comparing these techniques...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Ehrlichia canis; Dog; IFAT; Dot-ELISA; Nested PCR.
Ano: 2008 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782008000300027
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Clinical and epidemiological use of nested PCR targeting the repetitive element IS1111 associated with the transposase gene from Coxiella burnetii BJM
Mares-Guia,Maria Angélica M.M.; Guterres,Alexandro; Rozental,Tatiana; Ferreira,Michelle dos Santos; Lemos,Elba R.S..
ABSTRACT Q fever is a worldwide zoonosis caused by Coxiella burnetii—a small obligate intracellular Gram-negative bacterium found in a variety of animals. It is transmitted to humans by inhalation of contaminated aerosols from urine, feces, milk, amniotic fluid, placenta, abortion products, wool, and rarely by ingestion of raw milk from infected animals. Nested PCR can improve the sensitivity and specificity of testing while offering a suitable amplicon size for sequencing. Serial dilutions were performed tenfold to test the limit of detection, and the result was 10× detection of C. burnetti DNA with internal nested PCR primers relative to trans-PCR. Different biological samples were tested and identified only in nested PCR. This demonstrates the...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Q fever; Coxiella burnetii; Molecular diagnosis; Nested PCR; IS1111.
Ano: 2018 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822018000100138
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Detection of Mycoplasma hyopneumoniae in lungs and nasal swabs of pigs by nested PCR Arq. Bras. Med. Vet. Zootec.
Silva,F.M.F.; Castro,L.A.; Silva Júnior,A.; Moraes,M.P.; Moreira,M.A.S.; Almeida,M.R..
Fifty-four samples were collected from growing and finishing pigs for the molecular diagnosis of enzootic porcine pneumonia. Nineteen lung fragments were obtained from pigs that showed signs of respiratory disease and 35 nasal swabs were obtained from clinically healthy pigs. For the detection of the bacterial genome in the samples, the nested PCR technique was used to amplify a fragment of 706bp. This fragment was subsequently cloned and sequenced. The sequence of obtained nucleotides was compared with six other sequences of Mycoplasma hyopneumoniae and 11 sequences of other bacteria available in the Genbank. To measure the sensitivity of the nested PCR, serial dilutions (10-1 to 10-15) of cloned fragments were conducted based on the concentration of...
Tipo: Info:eu-repo/semantics/report Palavras-chave: Swine; Enzootic pneumonia; Mycoplasma hyopneumoniae; Nested PCR.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352009000100021
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Diagnóstico de Cryptosporidium spp. por meio de nested-PCR em caprinos jovens de Quixadá, Ceará. Repositório Alice
OLIVEIRA, D. A. dos S.; BRITO, R. L. L. de; INÁCIO, S. V.; OLIVEIRA, B. C. M.; BRESCIANI, K. D. S.; MEIRELES, M. V.; VIEIRA, L. da S..
2014
Tipo: Resumo em anais de congresso (ALICE) Palavras-chave: Nested PCR; Caprino; Cabrito; Diagnostico; Cryptosporidium; Goats; Kids; Diagnosis; PCR.
Ano: 2014 URL: http://www.alice.cnptia.embrapa.br/handle/doc/1007275
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Ehrlichia canis detection in dogs from Várzea Grande: a comparative analysis of blood and bone marrow samples Ciência Rural
Makino,Herica; Sousa,Valéria Régia Franco; Fujimori,Mahyumi; Rodrigues,Juliana Yuki; Dias,Alvaro Felipe Lima Ruy; Dutra,Valéria; Nakazato,Luciano; Almeida,Arleana do Bom Parto Ferreira de.
ABSTRACT: The objective of this study was to compare the DNA detection of Ehrlichia canis in blood and bone marrow to determine the prevalence of the agent in Várzea Grande, Mato Grosso. Blood samples and bone marrow from 80 dogs of both sexes, different breeds and age, were collected and processed for a cross-sectional study performed using nested PCR. Of the 80 dogs, 61 (76.3%) had E. canis DNA in one of the samples. The buffy coat was positive in 42 dogs (52.5%) and the bone marrow was positive in 33 (41.3%). There was no significant association between the positive biological samples of either the buffy coat or bone marrow and the presence or absence of clinical signs (P=0.49). No risk factor was associated with infection in the studied area. The bone...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Canine ehrlichiosis; Diagnosis; Nested PCR; Biological samples..
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782016000200310
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Importance of the association of molecular and immunological diagnosis in immunocompetent patient with Histoplasma capsulatum and Cryptoccocus neoformans infection: a case report J. Venom. Anim. Toxins incl. Trop. Dis.
Dantas,Kátia Cristina; Freitas,Roseli Santos de; Garcia,Roberta Scholz Pinto; Silva,Marcos Vinícius da; Muricy,Edna Cleide Mendes; Kohara,Valdelene Sayuri; Vicentini,Adriana Pardini.
This case reports an immunocompetent 29-year-old woman with suspected pneumonia, suggestive of fungal infection. Immunoblotting analysis reactivity againstHistoplasma capsulatum and Paracoccidioides brasiliensis were observed. Nested-PCR in blood employing species-specific primers was positive for H. capsulatum andCryptococcus neoformans. The evaluation of paucisymptomatic patients with positive results for H. capsulatum and C. neoformans could be relevant for the prevention as well as the possible evaluation of the reactivated quiescent foci. In conclusion, the associated methodology may have contributed to the monitoring endogenous reactivation of these diseases.
Tipo: Info:eu-repo/semantics/report Palavras-chave: Immunocompetent individuals; Nested PCR; Immunoblotting; Fungal infections; Blood fluid.
Ano: 2014 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992014000200507
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Multiple types of human papillomavirus in cervical samples in women in Campo Grande, MS, Brazil BJID
Tozetti,Inês Aparecida; Scapulatempo,Ilzia Doraci Lins; Kawski,Vicky Liege; Ferreira,Antonio Walter; Levi,José Eduardo.
We analyzed 87 cervical samples from Campo Grande, Mato Grosso do Sul, with a PGMY/GP+ nested PCR system. Positive samples were typed using E7 type-specific primer pairs for HPV 6/11, 16, 18, 45 and 66. Eighteen samples (22%) were infected with HPV6/11, 18 samples (22%) with HPV66, 13 samples (15.9%) with HPV45, 8 samples (9.8%) with HPV18 and 7 samples (8.5%) with HPV16. Seventeen samples (20.7%) were infected by two HPV types, and five samples (6.1%) by three HPV types. We conclude that infection with multiple types is present at a high frequency in our population and that there is a relation between some types and cytological finds.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Human papillomavirus; Multiple types; Nested PCR; TS-PCR.
Ano: 2006 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702006000500001
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Processamento de sêmen para extração de RNA genômico do vírus da artrite-encefalite caprina e diagnóstico molecular por RT-nested PCR. Repositório Alice
SIDER, L. H.; OLIVEIRA, A. N. de; VERAS, A. K. A.; KADRI, S. M.; FRANCO, M. M.; PINHEIRO, R. R.; ANDRIOLI, A..
O presente comunicado descreve os procedimentos necessários para a coleta e processamento de amostras de sêmen de caprinos infectados pelo CAEV para posterior extração do RNA viral por meio de um método baseado em centrifugação em coluna de sílica. A avaliação da presença de RNA no sêmen será feita, diretamente, por meio da reação de RT-nested PCR, portencial método de diagnóstico molecular da CAE.
Tipo: Outras publicações científicas (ALICE) Palavras-chave: Artrite-encefalite caprina; CAE; Caprino; Doença animal; Vírus; Diagnóstico; Diagnóstico; Goats; Animal diseases; Nested PCR; Caprine arthritis encephalit virus.
Ano: 2010 URL: http://www.alice.cnptia.embrapa.br/handle/doc/880100
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RpoB-PCR amplified gene and temporal temperature gradient gel electrophoresis: a rapid tool to analyse bacterial strains representative of cold-smoked salmon microflora ArchiMer
Giacomazzi, Sophie; Leroi, Francoise; L'Henaff, Cecile; Joffraud, Jean-jacques.
Aims: To evaluate rpoB gene as a biomarker of microbial biodiversity associated to cold-smoked salmon by a novel nested-polymerase chain reaction/temporal temperature gradient gel electrophoresis (PCR/TTGE) technique applied on pure cultures of reference strains. Methods and Results: DNA obtained from pure cultures of reference strains was used in a succession of a first PCR amplification of rpoB fragment with degenerated nonclamped primers and a nested-PCR with nondegenerated clamped primers. PCR products were then applied on a TTGE gel in order to analyse strains profile. High quantity of nested-PCR products were obtained for each tested strain and TTGE profiles showed a good separation between the different reference bacteria and an easy way to...
Tipo: Text Palavras-chave: TTGE; RpoB; Nested PCR; Cold smoked salmon flora.
Ano: 2004 URL: http://archimer.ifremer.fr/doc/2004/publication-579.pdf
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Soroconversão e controle de infecção do lentivírus caprino. Repositório Alice
ALVES, R. P. A..
Resumo: O vírus da Artrite Encefalite Caprina (CAEV) pertence à família Retroviridae, subfamília Lentivirinae, gênero Lentivírus. Ele causa uma infecção denominada de Artrite Encefalite Caprina (AEC) que é capaz de se disseminar em um rebanho por um longo período. Com o presente estudo objetivou-se acompanhar a soroconversão da AEC, utilizando teste sorológico (Western Blotting), e a dinâmica de detecção da doença, com o uso da técnica molecular Reação em Cadeia de Polimerase nested (PCR nested). Dois lotes distintos de animais foram utilizados em momentos diferentes, sendo o primeiro formado com cabritos da raça Saanen, composto por 47 animais, e o segundo formado por 24 cabritos das raças Saanen e Anglo-nubiana. Em ambos os lotes realizaram-se testes de...
Tipo: Tese/dissertação (ALICE) Palavras-chave: Nested PCR; PCR nested; Artrite Encefalite Caprina; CAE; CAEV; Immunological techniques; Caprino; Ovino; Doença Animal; Erradicação; Small ruminants; Sheep diseases; Goats; Goat diseases; Western blotting; Lentivirus; Caprine arthritis encephalitis virus; Immunoblotting; Genetic techniques and protocols; Disease control; Brazil.
Ano: 2019 URL: http://www.alice.cnptia.embrapa.br/alice/handle/doc/1119491
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Species-specific nested PCR as a diagnostic tool for Brucella ovis infection in rams Arq. Bras. Med. Vet. Zootec.
Costa,L.F.; Nozaki,C.N.; Lira,N.S.C.; Antunes,J.M.A.P.; Xavier,M.N.; Costa,E.A.; Paixão,T.A.; Santos,R.L.; Megid,J..
The aim of the present study was to evaluate a species-specific nested PCR based on a previously described species-specific PCR for detection of B. ovis in semen and urine samples of experimentally infected rams. The performance of the species-specific nested PCR was compared with the results of a genus-specific PCR. Fourteen rams were experimentally infected with the Brucella ovis REO 198 strain and samples of semen and urine were collected every week up to 180 days post infection. Out of 83 semen samples collected, 42 (50.6%) were positive for the species-specific nested PCR, and 23 (27.7%) were positive for the genus-specific PCR. Out of 75 urine samples, 49 (65.3%) were positive for the species-specific nested PCR, whereas 11 (14.6%) were...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Brucella ovis; Species-specific; Nested PCR; Semen; Urine.
Ano: 2013 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352013000100009
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Uso da técnica de swim-up para a remoção do Vírus da Artrite Encefalite Caprina do sêmen de reprodutores infectados. Repositório Alice
ÁVILA, A. A.; SIDER, L. H.; VERAS, A. K. A.; PINHEIRO, R. R.; OLIVEIRA, M. L. M.; SILVA, P. A. F.; SOUSA, S. D.; ANDRIOLI, A..
Resumo: Neste estudo, 67 ejaculados foram avaliados, antes e depois da técnica de swim-up, em relação à qualidade seminal e à presença do CAEV. Das 67 amostras testadas por PCRn, antes do swim-up, 47 (70,15%) foram positivas para o DNA pró-viral. No entanto, quatro amostras adicionais foram positivas ao RT-nested PCR após o swim-up, o que permite dizer que, pelo menos, 76,12% (51/67) delas estavam infectadas antes da lavagem. Todavia, em 23,88% (16/67) das amostras não foi detectada a presença do CAEV. Após a aplicação da técnica de swim-up, constatou-se, pela PCRn e RT-nested PCR, que houve uma redução significativa (X²= 9,078; p<0,001) da presença do CAEV nas amostras seminais, pois 28 de 51 amostras positivas resultaram livres do vírus (54,90%),...
Tipo: Artigo em periódico indexado (ALICE) Palavras-chave: CAEV; PCRn; Lavagem seminal; Artrite Encefalite Caprina; Seminal washing; RT-nested PCR; Nested PCR; RT-nested PCR; Percoll; Caprino; Sêmen; Goats; DNA.
Ano: 2015 URL: http://www.alice.cnptia.embrapa.br/handle/doc/1032730
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Uso da técnica de swim-up para a remoção do vírus da artrite encefalite caprina e obtenção de espermatozoides viáveis. Repositório Alice
ÁVILA, A. A..
Os procedimentos de lavagem seminal associados a testes moleculares de alta sensibilidade para o diagnóstico do vírus da Artrite Encefalite Caprina (CAEV), como a Nested PCR e a RT-nested PCR, podem ter um impacto significante na prevenção da transmissão de enfermidades e preservação do germoplasma de reprodutores portadores do CAEV, a fim de ser utilizado sem o risco de contaminação da fêmea por via reprodutiva. Nesse cenário, objetivou-se com o presente trabalho determinar a influência da técnica de swim-up sobre a viabilidade espermática e sua eficiência na remoção de partículas virais de amostras de sêmen de reprodutores caprinos portadores do CAEV. Nesse estudo, 67 ejaculados foram avaliados antes da técnica de swim-up, quanto ao volume, concentração,...
Tipo: Tese/dissertação (ALICE) Palavras-chave: Nested PCR; RT-nested PCR; CAEV; Sperm selection; Semen quality; Seminal washing; Artrite-encefalite caprina; Swim-up; Caprino; Doença animal; Vírus; Sêmen; Espermatozóide; Fertility; Goats; Caprine arthritis encephalit virus; Spermatozoa; Lentivirus; Movement.
Ano: 2013 URL: http://www.alice.cnptia.embrapa.br/handle/doc/997345
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Validation and clinical application of a molecular method for the identification of Cryptococcus neoformans/Cryptococcus gattii complex DNA in human clinical specimens BJID
Rivera,Vanessa; Gaviria,Marcela; Muñoz-Cadavid,Cesar; Cano,Luz; Naranjo,Tonny.
ABSTRACT The diagnosis of cryptococcosis is usually performed based on cultures of tissue or body fluids and isolation of the fungus, but this method may require several days. Direct microscopic examination, although rapid, is relatively insensitive. Biochemical and immunodiagnostic rapid tests are also used. However, all of these methods have limitations that may hinder final diagnosis. The increasing incidence of fungal infections has focused attention on tools for rapid and accurate diagnosis using molecular biological techniques. Currently, PCR-based methods, particularly nested, multiplex and real-time PCR, provide both high sensitivity and specificity. In the present study, we evaluated a nested PCR targeting the gene encoding the ITS-1 and ITS-2...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cryptococcus neoformans/Cryptococcus gattiicomplex; Nested PCR; Cryptococcosis; Molecular diagnosis.
Ano: 2015 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702015000600563
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Validation and clinical application of a nested PCR for paracoccidioidomycosis diagnosis in clinical samples from Colombian patients BJID
Gaviria,Marcela; Rivera,Vanessa; Muñoz-Cadavid,Cesar; Cano,Luz Elena; Naranjo,Tonny Williams.
Paracoccidioidomycosis is a systemic and endemic mycosis, restricted to tropical and subtropical areas of Latin America. The infection is caused by the thermal dimorphic fungus Paracoccidioides brasiliensisand Paracoccidioides lutzii. The diagnosis of paracoccidioidomycosis is usually performed by microscopic examination, culture and immunodiagnostic tests to respiratory specimens, body fluids and/or biopsies; however these methods require laboratory personnel with experience and several days to produce a result. In the present study, we have validated and evaluated a nested PCR assay targeting the gene encoding the Paracoccidioides gp43membrane protein in 191 clinical samples: 115 samples from patients with proven infections other than...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Paracoccidioides brasiliensis; Nested PCR; Paracoccidioidomycosis; Molecular diagnosis.
Ano: 2015 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702015000400376
Registros recuperados: 15
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