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Perfil de restrição de um fragmento do gene da hemaglutinina amplificado pela RT-PCR a partir de estirpes vacinais e selvagens do vírus da cinomose canina Arq. Bras. Med. Vet. Zootec.
Negrão,F.J.; Wosiacki,S.H.; Alfieri,A.A.; Alfieri,A.F..
Fragmentos com 721 pares de bases do gene da hemaglutinina (H) do vírus da cinomose canina (CDV), amplificados pela RT-PCR a partir de três estirpes vacinais (Snyder Hill, Onderstepoort e Rockborn) e de 27 amostras de campo, provenientes de cães com cinomose, foram clivados com as endonucleases Hinf I and Rsa I. A seleção das enzimas foi realizada por meio de análises in silico de seqüências do CDV depositadas em bases públicas de dados. Tanto as estirpes vacinais quanto as amostras selvagens do CDV apresentaram com a enzima Hinf I o mesmo perfil de restrição, confirmando a identidade do fragmento amplificado pela RT-PCR, uma vez que todas as estirpes com seqüências disponíveis (GenBank) têm sítios de restrição para essa enzima nas mesmas posições. O...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cão; Cinomose; Vírus da cinomose canina; Gene H; RT-PCR; RFLP.
Ano: 2006 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352006000600019
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PPM1B and P-IKKβ expression levels correlated inversely with rat gastrocnemius atrophy after denervation BJMBR
Wei,Jian; Liang,Bing-Sheng.
Activated inhibitor of nuclear factor-κB kinase β (IKKβ) is necessary and sufficient for denervated skeletal muscle atrophy. Although several studies have shown that Mg2+/Mn2+-dependent protein phosphatase 1B (PPM1B) inactivated IKKβ, few studies have investigated the role of PPM1B in denervated skeletal muscle. In this study, we aim to explore the expression and significance of PPM1B and phosphorylated IKKβ (P-IKKβ) during atrophy of the denervated gastrocnemius. Thirty young adult female Wistar rats were subjected to right sciatic nerve transection and were sacrificed at 0 (control), 2, 7, 14, and 28 days after denervation surgery. The gastrocnemius was removed from both the denervated and the contralateral limb. The muscle wet weight ratio was...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Muscular atrophy; Denervation; PPM1B; P-IKKβ; IKKβ; RT-PCR; Western blot.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2012000800004
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Prevalence of hepatitis A virus in sea food in Iran Arq. Bras. Med. Vet. Zootec.
Momtaz,H.; Rahimi,E.; Hajian,S.; Momeni,M..
The objective of this study was to determine the prevalence of Hepatitis A Virus (HAV) in sea food samples in the Isfahan and Shahrekord townships in Iran. From September 2010 to April 2011, a total of 300 samples of fresh fish, shrimp, crab and lobster were obtained from randomly selected retail stores in the Isfahan and Shahrekord townships in Iran. The samples were tested for the presence of HAV using a reverse transcriptase- polymerase chain reaction method. Out of the total number of samples examined, 8 (2.7%) were found to be positive for HAV. This virus was detected in 5% and 1.7% of fresh fish and shrimp, respectively. This study shows the importance of sea food as potential sources of HAV infection in people in Iran.
Tipo: Info:eu-repo/semantics/article Palavras-chave: HAV; Sea food; RT-PCR; Iran.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352012000400015
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Primer reporte de Tobacco mosaic virus (TMV) y Poinsettia mosaic virus (PnMV) en nochebuena de sol (Euphorbia pulcherrima Willd. Ex Klotzch) en México Phyton
Ocampo Ocampo,T; Ochoa Martínez,DL; Ramírez Rojas,S; Valdovinos Ponce,G; Nava Díaz,C.
El Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias (INIFAP), a través del Campo Experimental "Zacatepec" en el estado Morelos, inició un programa de mejoramiento genético de plantas silvestres y semicultivadas de nochebuena ("nochebuena de sol"). Dicho programa requiere de la generación de una base de datos fitosanitarios con tolerancia a los principales patógenos. México no cuenta con información sobre las enfermedades causadas por virus en la nochebuena de sol, por lo que el objetivo de la presente investigación fue generar datos preliminares sobre los virus asociados a estos materiales. Con base en los resultados obtenidos en las técnicas DAS-ELISA y RT-PCR, se reportan por primera vez en México los virus Tobacco mosaic virus...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Nochebuena silvestre; Nochebuena semicultivada; Virus; DAS-ELISA; RT-PCR.
Ano: 2013 URL: http://www.scielo.org.ar/scielo.php?script=sci_arttext&pid=S1851-56572013000200011
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Rabies virus in a pregnant naturally infected southern yellow bat (Lasiurus ega) J. Venom. Anim. Toxins incl. Trop. Dis.
Allendorf,SD; Albas,A; Cipriano,JRB; Antunes,JMAP; Appolinário,CM; Peres,MG; da Rosa,AR; Sodré,MM; Megid,J.
Current knowledge on bat lyssavirus infections in their native hosts is limited and little is known about the virulence, virus dissemination and transmission among free-living insectivorous bats. The present study is a brief description of rabies virus (RABV) dissemination in tissues of a naturally infected pregnant southern yellow bat (Lasiurus ega) and its fetuses, obtained by reverse-transcriptase polymerase chain reaction (RT-PCR). The RT-PCR was positive in samples from the brain, salivary gland, tongue, lungs, heart, kidneys and liver. On the other hand, the placenta, three fetuses, spleen, intestine and brown fat tissue tested negative. This research demonstrated the absence of rabies virus in the fetuses, thus, in this specific case, the...
Tipo: Info:eu-repo/semantics/other Palavras-chave: Rabies; Bats; Vertical transmission; RT-PCR.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992011000200014
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RT-PCR em pools de soros sangüíneos para o diagnóstico da infecção aguda e de animais persistentemente infectados pelo vírus da diarréia viral bovina Arq. Bras. Med. Vet. Zootec.
Pilz,D.; Alfieri,A.F.; Lunardi,M.; Alfieri,A.A..
Utilizou-se a técnica da RT-PCR para a detecção da região 5' UTR do genoma do vírus da diarréia viral bovina (BVDV) em pools de soros sangüíneos provenientes de um rebanho, constituído por 226 animais, que apresentava distúrbios da reprodução. A partir das amostras individuais de soro e de acordo com a categoria dos animais e o número de animais por categoria foram formados 10 pools (A a J) de soros. A primeira avaliação revelou a amplificação de um produto com 290pb nas reações referentes aos grupos D (35 vacas) e H (25 bezerros lactentes) que, após o desmembramento em amostras individuais, resultou na identificação de 11 vacas lactantes e 12 bezerros em amamentação positivos. Para a identificação de animais persistentemente infectados (PI) entre os 23...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Bovino; Diarréia viral bovina; BVDV; RT-PCR.
Ano: 2007 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352007000100001
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RT-PCR for confirmation of echovirus 30 isolated in Belém, Brazil BJID
Gomes,Maria de Lourdes C.; Ferreira,Lauze Lee A.; Gomes,Rodrigo Henryque; Lamarão,Letícia M.; Silveira,Edna da; Rodrigues,Lílian da Silva; Silva,Cláudio Ferreira da; Almeida,Eduardo Ribeiro de.
Echovirus (Echo) 30 or human enterovirus B is the most frequent enterovirus associated with meningitis cases. Epidemics and outbreaks of this disease caused by Echo 30 have occurred in several countries. In Brazil, Echo 30 has been isolated from sporadic cases and outbreaks that occurred mainly in the south and southeast regions. We used RT-PCR to examine Echo 30 isolates from meningitis cases detected from March 2002 to December 2003 in Belém, state of Pará, in northern Brazil. The patients were attended in a Basic Health Unit (State Health Secretary of Pará), where cerebrospinal fluid (CSF) was collected and stored in liquid nitrogen. Weekly visits were made by technicians from Evandro Chagas Institute to the health unit and samples were stored at -70ºC...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Echovirus 30; Aseptic meningitis; RT-PCR; Belém; Brazil.
Ano: 2007 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702007000400006
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Secuenciación de un fragmento de ADNc homólogo a interleucina-1 alfa humana derivado de leucocitos de armadillo (Dasypus novemcinctus) Veterinaria México
Flores-Medina,Saúl; Díaz-García,Francisco J.; Guerra-Infante,Fernando M..
Se realizó detección del ARN mensajero (ARNm) mediante un ensayo de transcripción reversa acoplada a la reacción en cadena de la polimerasa (RT-PCR) específico para la detección de Interleucina-1 alfa humana en leucocitos de armadillo estimulados con acetato de forbol miristato (PMA, por su siglas en inglés); esta estrategia permitió amplificar un fragmento de ADN de 491 pares de bases. Adicionalmente, la secuencia mostró alta homología nucleotídica con las secuencias de ADNc humano (99%), mono (93%), cerdo (82%), caballo (81%) y llama (81%), mientras que la secuencia deducida de aminoácidos fue compatible con el precursor de la proteína IL-1a humana. Estos resultados sugieren presencia del gen de interleucina-1 en el genoma del armadillo; sin embargo, es...
Tipo: Info:eu-repo/semantics/report Palavras-chave: Dasypus novemcinctus; Interleucina-1; RT-PCR; Secuenciación.
Ano: 2008 URL: http://www.scielo.org.mx/scielo.php?script=sci_arttext&pid=S0301-50922008000300008
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SECUENCIACIÓN DEL GENOMA COMPLETO DEL Potato yellow vein virus (PYVV) EN TOMATE (Solanum lycopersicum) EN COLOMBIA Acta biol.Colomb.
MUÑOZ BAENA,Laura; GUTIÉRREZ SÁNCHEZ,Pablo Andrés; MARÍN MONTOYA,Mauricio.
RESUMEN Potato yellow vein virus (PYVV), es uno de los fitopatógenos más limitantes para la producción de papa en la región de Los Andes. A pesar que se le ha detectado infectando tomate en Colombia, el conocimiento de las características biológicas de las cepas presentes en este hospedante es muy limitado. En este estudio, utilizando secuenciación masiva de nueva generación (NGS), se obtuvo la secuencia completa de los tres segmentos genómicos del PYVV en plantas de tomate en Marinilla (Antioquia) y se evalúo la utilidad de tres juegos de cebadores para su detección mediante pruebas de RT-PCR convencional y en tiempo real (RT-qPCR). El genoma de la secuencia consenso presentó tamaños de 8043 nt (ARN1), 5346 nt (ARN2) y 3896 nt (ARN3) y se identificaron...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Crinivirus; RT-PCR; RT-qPCR; Secuenciación de nucleótidos de alto rendimiento; Solanaceae.
Ano: 2017 URL: http://www.scielo.org.co/scielo.php?script=sci_arttext&pid=S0120-548X2017000100001
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Sensitivity of RT-PCR method in samples shown to be positive for Zika virus by RT-qPCR in vector competence studies Genet. Mol. Biol.
Paiva,Marcelo Henrique Santos; Guedes,Duschinka Ribeiro Duarte; Leal,Walter Soares; Ayres,Constância Flávia Junqueira.
Abstract Tissue samples from mosquitoes artificially infected with Zika virus and shown to be positive by RT-qPCR were reexamined by RT-PCR. Using these samples we compared the two methods employed in virus RNA detection for vector competence studies. Results demonstrated that, albeit useful, RT-PCR gave false negatives with low viral loads (< 106 RNA copies/ml).
Tipo: Info:eu-repo/semantics/article Palavras-chave: ZIKV; RT-PCR; Aedes aegypti; Vector competence.
Ano: 2017 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572017000400597
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Sequence analysis of the capsid protein gene of an isolate of Apple stem grooving virus, and its survey in Southern Brazil Trop. Plant Pathol.
NICKEL,OSMAR; FAJARDO,THOR V.M.; JELKMANN,WILHELM; KUHN,GILMAR B..
Apple stem grooving virus (ASGV) is one of the most important viruses infecting fruit trees. This study aimed at the molecular characterization of ASGV infecting apple (Malus domestica) plants in Santa Catarina (SC). RNA extracted from plants infected with isolate UV01 was used as a template for RT-PCR using specific primers. An amplified DNA fragment of 755 bp was sequenced. The coat protein gene of ASGV isolate UV01 contains 714 nucleotides, coding for a protein of 237 amino acids with a predicted Mr of approximately 27 kDa. The nucleotide and the deduced amino acid sequences of the coat protein gene showed identities of 90.9% and 97.9%, respectively, with a Japanese isolate of ASGV. Very high amino acid homologies (98.7%) were also found with Citrus...
Tipo: Info:eu-repo/semantics/other Palavras-chave: Molecular characterization; Capillovirus; RT-PCR.
Ano: 2001 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-41582001000300014
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Sequencing and phylogenetic analysis of the infectious bursal disease virus isolates from outbreak in layer flocks in the state of Minas Gerais BJM
Dias,Camila Cristina Almeida; Souza,Fernanda de Oliveira; Silva,Elisa Monteiro Sant'Anna da; Eller,Monique Renon; Barrios,Priscilla Rochele; Santos,Bernadete Miranda dos; Moraes,Mauro Pires; Almeida,Márcia Rogéria de.
Sequencing and phylogenetic analysis based on the nucleotide sequence of the gene encoding VP2 protein was carried out in order to characterize the agent of two outbreaks of infectious bursal disease in layer flocks in the state of Minas Gerais in 2004. The results indicate the outbreaks could be related to the vaccinal virus.
Tipo: Info:eu-repo/semantics/article Palavras-chave: IBDV; RT-PCR; Molecular characterization; Phylogenic analysis; Infectious bursal disease.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822009000100036
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Sm14 gene expression in different stages of the Schistosoma mansoni life cycle and immunolocalization of the Sm14 protein within the adult worm BJMBR
Brito,C.F.A.; Oliveira,G.C.; Oliveira,S.C.; Street,M.; Riengrojpitak,S.; Wilson,R.A.; Simpson,A.J.G.; Correa-Oliveira,R..
Sm14 is a 14-kDa vaccine candidate antigen from Schistosoma mansoni that seems to be involved in cytoplasmic trafficking of fatty acids. Although schistosomes have a high requirement for lipids, they are not able to synthesize fatty acids and sterols de novo. Thus, they must acquire host lipids. In order to determine whether Sm14 is present in different stages of the life cycle of the parasite, we performed RT-PCR. Sm14 mRNA was identified in all stages of the life cycle studied, mainly schistosomulum, adult worm and egg. Additionally, we used a rabbit anti-Sm14 polyclonal antibody in an indirect immunofluorescence assay to localize Sm14 in adult worm sections. The basal lamella of the tegument and the gut epithelium were strongly labeled. These tissues...
Tipo: Info:eu-repo/semantics/other Palavras-chave: Sm14; Fatty acid-binding protein; FABP; Immunolocalization; RT-PCR.
Ano: 2002 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2002000300014
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Tomato chlorotic spot virus in hydroponically-grown lettuce in São Paulo State, Brazil Trop. Plant Pathol.
Colariccio,Addolorata; Eiras,Marcelo; Chaves,Alexandre L. R.; Harakava,Ricardo; Chagas,César M..
In the regions of Campinas and Sumaré, São Paulo, Brazil, hidroponically grown crops of Lettuce (Lactuca sativa) cv. Verônica, which showed virus-like symptoms were examined by electron microscope, biological, serological and molecular tests. Pleomorphic, enveloped particles (80-100 nm in diameter) were always detected in these samples. Experimentally inoculated host plants, including lettuce, reacted with tospoviruses-induced symptoms. Some differences were observed in Gomphrena globosa, which reacted by showing local lesions and systemic mosaic. Two isolates of Tomato chlorotic spot virus (TCSV) were identified by DAS-ELISA and by RT-PCR. The sequencing and alignment of the RT-PCR coat protein amplified fragments have indicated a high degree of homology...
Tipo: Info:eu-repo/semantics/other Palavras-chave: Tospovirus; Serology; RT-PCR; Sequencing.
Ano: 2004 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-41582004000300012
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TRANSCRIPTOS DE FUSIÓN DEL GEN BCR/ABL EN PACIENTES CON LEUCEMIA MIELOIDE CRÓNICA International Journal of Morphology
Artigas,Carmen Gloria; Melo,Angélica; Roa,Juan Carlos; Roa,Iván; Quijada,Ingrid; Vittini,Cecilia; Cabrera,María Elena; Risueño,Concepción.
La anormalidad citogenética más común en la leucemia mieloide crónica (LMC) es el cromosoma Philadelphia, producida por la t(9;22), cuya expresión molecular es el gen de fusión BCR-ABL, que codifica proteínas con actividad tirosinquinasa. Según el punto de ruptura de los genes BCR o ABL se produce una proteína de fusión de 210-kD(p210) o 190-kD(p190). La presencia de este gen de fusión en pacientes con LMC tiene implicancia diagnóstica. Con el propósito de detectar transcriptos de fusión del gen BCR/ABL en pacientes con leucemia mieloide crónica, procedentes de la IX Región de Chile, se estudiaron 14 muestras de sangre de 11 pacientes con LMC. A 2 de ellos, se les realizó seguimiento durante su tratamiento con Gleevec. Se aplicó la técnica de reacción en...
Tipo: Journal article Palavras-chave: BCR/ABL; RT-PCR; Leucemia mieloide crónica; Chile.
Ano: 2003 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-95022003000300004
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Up-regulated expression of CD147 gene in malignant bone tumor and the possible induction mechanism during osteoclast formation BJMBR
Qiao,Suchi; Liu,Chang; Xu,Weijie; AZhaTi,WuBuLi; Li,Cheng; Wang,Zhiwei.
It is increasingly evident that the microenvironment of bone can influence cancer phenotype in many ways that favor growth in bone. CD147, a transmembrane protein of the immunoglobulin (Ig) superfamily, was identified independently in different species and has many designations across different species. However, expression levels of CD147 mRNA in bone cancer have not been described. In this study, we have used real-time fluorescence quantification (RT-PCR) to demonstrate CD147 expression in malignant bone cancer and benign bone tumor tissues. The results suggested that the expression of CD147 gene was significantly up-regulated in malignant bone cancer. Moreover, we found that over-expressed RANKL progressively enhanced osteoclast formation up to 48 h,...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Bone cancer; RT-PCR; Western blot; RANKL; Human.
Ano: 2018 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2018000900608
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Variabilidade genética de Sugarcane mosaic virus, causando mosaico em milho no Brasil PAB
Gonçalves,Marcos Cesar; Galdeano,Diogo Manzano; Maia,Ivan de Godoy; Chagas,César Martins.
O objetivo deste trabalho foi caracterizar biológica e molecularmente três isolados de Sugarcane mosaic virus (SCMV) de lavouras de milho, analisá-los filogeneticamente e discriminar polimorfismos do genoma. Plantas com sintomas de mosaico e nanismo foram coletadas em lavouras de milho, no Estado de São Paulo e no Município de Rio Verde, GO, e seus extratos foliares foram inoculados em plantas indicadoras e submetidos à análise sorológica com antissoros contra o SCMV, contra o Maize dwarf mosaic virus (MDMV) e contra o Johnsongrass mosaic virus (JGMV). Mudas de sorgo 'Rio' e 'TX 2786' apresentaram sintomas de mosaico após a inoculação dos três isolados, e o DAS-ELISA confirmou a infecção pelo SCMV. O RNA total foi extraído e usado para amplificação por...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Potyviridae; Zea mays; Mosaico comum do milho; RFLP; RT-PCR.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-204X2011000400004
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Virus de Distemper Canino: detección molecular de diferentes aislamientos provenientes de perros de la provincia de santa fe, argentina, entre los años 2000 y 2010 InVet
Pinotti,M.; Gollan,A; Canavesio,M.; Passeggi,C.; Larrateguy,M.V.; Paz,M.E.; Formentini,E..
En la ciudad de Santa Fe, Argentina, durante los años 2000 y 2010, se aislaron 13 cepas autóctonas del virus Distemper Canino. Las muestras se tomaron de caninos con signos clínicos de la enfermedad y consistieron en hisopados nasales, oculares, faríngeos (tonsilas) y conjuntivales. Los aislamientos se realizaron en células MDCK, las cuales mostraron efecto citopatogénico característico. Los virus provenientes de la cosecha, se detectaron por RT-PCR de una región que codifica la nucleoproteína NP asociada al RNA (posición 769 a la 1055 del genoma) obteniéndose en todos los casos un producto de 287 pb comparable con el obtenido con cepas vacunales (cepas Ondersterpoort y Lederle), lo que indica presencia viral en todas las muestras procesadas. Las cepas...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Virus; Distemper canino; Estudio molecula; RT-PCR.
Ano: 2016 URL: http://www.scielo.org.ar/scielo.php?script=sci_arttext&pid=S1668-34982016000200007
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Virus isolation and genotype identification of human respiratory syncytial virus in Guizhou Province, China BJID
Zhao,Ting; Ye,Zhixu; Wang,Binlin; Cui,Yuxia; Nie,Yingjie; Yang,Bin; Chen,Kun; Zhang,Hua; Hu,Fangfang; Yu,Fuxun.
ABSTRACT To investigate the genetic variation and molecular epidemiology characteristics of Human Respiratory Syncytial Virus (HRSV) in Guizhou Province, nasopharyngeal aspirates were collected from patients with acute respiratory infection (ARI) in Guizhou Provincial People's Hospital, from December 2017 to March 2018, and inoculated to Hep-2 cells to isolate HRSV. Cells that showed cytopathic effect (CPE) were then confirmed by indirect immunofluorescence assay and reverse transcription. The sequence of the PCR products was determined for HRSV isolates, and the genetic variation was analyzed. Out of 196 nasopharyngeal aspirate samples, HRSV were isolated in 39. The second hypervariable region at the 3' terminal of glycoprotein gene (HVR2) sequence...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Human respiratory syncytial virus; Virus isolation; RT-PCR; HVR2; Phylogenetic analysis; Molecular epidemiology.
Ano: 2019 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702019000600427
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Virus isolation and molecular characterization of canine distemper virus by RT-PCR from a mature dog with multifocal encephalomyelit BJM
Amude,Alexandre Mendes; Carvalho,Glei dos Anjos; Alfieri,Amauri Alcindo; Alfieri,Alice Fernandes.
A case of multifocal distemper encephalomyelitis in a mature dog is described. In the presented case the ante mortem clinical diagnosis of canine distemper virus (CDV) infection could not be ideally performed due to the absence of typical signs of distemper, such as myoclonus and systemic signs accompanying the nervous signs. The definitive diagnosis of distemper encephalomyelitis was only carried out at post mortem through virus isolation in cell culture from fresh central nervous system (CNS) fragments and CDV nucleoprotein gene detection in the CNS by RT-PCR.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Dog; Canine distemper virus; Encephalomyelitis; RT-PCR.
Ano: 2007 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822007000200031
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