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A Rapid and Efficient Method for Purifying High Quality Total RNA from Peaches (Prunus persica) for Functional Genomics Analyses Biol. Res.
MEISEL,LEE; FONSECA,BEATRIZ; GONZÁLEZ,SUSANA; BAEZA-YATES,RICARDO; CAMBIAZO,VERONICA; CAMPOS,REINALDO; GONZALEZ,MAURICIO; ORELLANA,ARIEL; RETAMALES,JULIO; SILVA,HERMAN.
Prunus persica has been proposed as a genomic model for deciduous trees and the Rosaceae family. Optimized protocols for RNA isolation are necessary to further advance studies in this model species such that functional genomics analyses may be performed. Here we present an optimized protocol to rapidly and efficiently purify high quality total RNA from peach fruits (Prunus persica). Isolating high-quality RNA from fruit tissue is often difficult due to large quantities of polysaccharides and polyphenolic compounds that accumulate in this tissue and co-purify with the RNA. Here we demonstrate that a modified version of the method used to isolate RNA from pine trees and the woody plant Cinnamomun tenuipilum is ideal for isolating high quality RNA from the...
Tipo: Journal article Palavras-chave: CDNA library; Fruit; Fruit trees; Functional genomics; Peach; Polysaccharides; Prunus persica; RNA isolation; Rosaceae; RT-PCR.
Ano: 2005 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602005000100010
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Acute promyelocytic leukemia: the study of t(15;17) translocation by fluorescent in situ hybridization, reverse transcriptase-polymerase chain reaction and cytogenetic techniques BJMBR
Chauffaille,M.L.L.F.; Figueiredo,M.S.; Beltrani,R.; Antunes,S.V.; Yamamoto,M.; Kerbauy,J..
Acute promyelocytic leukemia (AML M3) is a well-defined subtype of leukemia with specific and peculiar characteristics. Immediate identification of t(15;17) or the PML/RARA gene rearrangement is fundamental for treatment. The objective of the present study was to compare fluorescent in situ hybridization (FISH), reverse transcriptase-polymerase chain reaction (RT-PCR) and karyotyping in 18 samples (12 at diagnosis and 6 after treatment) from 13 AML M3 patients. Bone marrow samples were submitted to karyotype G-banding, FISH and RT-PCR. At diagnosis, cytogenetics was successful in 10 of 12 samples, 8 with t(15;17) and 2 without. FISH was positive in 11/12 cases (one had no cells for analysis) and positivity varied from 25 to 93% (mean: 56%). RT-PCR was done...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Acute promyelocytic leukemia; Karyotype; FISH; RT-PCR; PML/RARA genem rearrangement.
Ano: 2001 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2001000600006
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Ajustes em protocolo de multiplex voltado à detecção de vírus de videira presentes no Submédio do Vale do São Francisco. Repositório Alice
SILVA, A. dos A.; FREITAS, D. M. S..
Neste trabalho, testou-se diversos ajustes para um RT-PCR multiplex, que utiliza diferentes pares de oligonucleotídeos ao mesmo tempo, o que possibilita a detecção simultânea de várias espécies de vírus, fator que diminui o tempo e o custo das análises.
Tipo: Parte de livro Palavras-chave: Vale do Sao Francisco; RT-PCR; Uva; Vírus; Doença; Grapes; Vitis.
Ano: 2018 URL: http://www.alice.cnptia.embrapa.br/alice/handle/doc/1099008
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An isolate of Apple stem grooving virus associated with Cleopatra mandarin fruit intumescence Trop. Plant Pathol.
Lovisolo,Osvaldo; Accotto,Gian Paolo; Masenga,Vera; Colariccio,Addolorata.
A citrus tatter leaf isolate (CTLV-Cl) of Apple stem grooving virus (ASGV) has been found to be associated with a fruit rind intumescence in Cleopatra mandarin (Citrus reshni) in Limeira (SP). The CTLV-Cl was mechanically transmitted to the main experimental herbaceous hosts of CTLV. Chenopodium quinoa and C. amaranticolor reacted with local lesions and systemic symptoms while other test plants reacted somewhat differently than what is reported for CTLV. A pair of primers designed for specific detection of ASGV and CTLV amplified the expected 801 bp fragment from the CTLV-Cl-infected plants. Typical capillovirus-like particles were observed by the electron microscope in experimentally infected C. quinoa and C. amaranticolor leaves.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Citrus tatter leaf virus; Capillovirus; RT-PCR.
Ano: 2003 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-41582003000100008
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Anoxia- and hypoxia-induced expression of LDH-A* in the Amazon Oscar, Astronotus crassipinis Genet. Mol. Biol.
Almeida-Val,Vera Maria Fonseca; Oliveira,Alice Reis; Silva,Maria de Nazaré Paula da; Ferreira-Nozawa,Monica Stropa; Araújo,Roziete Mendes; Val,Adalberto Luis; Nozawa,Sérgio Ricardo.
Adaptation or acclimation to hypoxia occurs via the modulation of physiologically relevant genes, such as erythropoietin, transferrin, vascular endothelial growth factor, phosphofructokinase and lactate dehydrogenase A. In the present study, we have cloned, sequenced and examined the modulation of the LDH-A gene after an Amazonian fish species, Astronotus crassipinis (the Oscar), was exposed to hypoxia and anoxia. In earlier studies, we have discovered that adults of this species are extremely tolerant to hypoxia and anoxia, while the juveniles are less tolerant. Exposure of juveniles to acute hypoxia and anoxia resulted in increased LDH-A gene expression in skeletal and cardiac muscles. When exposed to graded hypoxia juveniles show decreased LDH-A...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Amazon; Fishes; Gene expression; RT-PCR; LDH-A; Hypoxia; Anoxia.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572011000200025
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Apoptose e expressão de VP2 e GAPDH na infecção precoce pelo vírus da doença infecciosa da bursa de Fabricius em pintos SPF Arq. Bras. Med. Vet. Zootec.
Batista,J.J.; Martins,A.S.; Moro,L.; Resende,J.S.; Martins,N.R.S.; Vasconcelos,A.C..
Vinte e nove pintos SPF de um dia foram inoculados com o vírus da doença infecciosa da bursa de Fabricius (VDIB) para avaliar a ocorrência precoce de apoptose e a expressão da proteína viral 2 (VP2) e da enzima gliceraldeído fosfato dehidrogenase (GAPDH). Os animais foram distribuídos em cinco grupos: 1-controle; e 2 a 5- com 24, 48, 72 e 96 horas pós-inoculação, respectivamente. Fragmentos da bursa de Fabricius foram colhidos para processamento histológico e extração de RNA. Lâminas coradas em HE e TUNEL (marcação in situ da fragmentação do genoma com transferase terminal de deoxinucleotídeo) foram utilizadas na morfometria do índice apoptótico. Amostras de mRNA foram testadas para a expressão dos genes VP2 e GAPDH utilizando-se transcrição reversa e...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Apoptose; IBDV; Bursa de Fabricius; RT-PCR.
Ano: 2007 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352007000200007
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Assessment of cytokine values in serum by RT-PCR in HIV-1 infected individuals with and without highly active anti-retroviral therapy (HAART) J. Venom. Anim. Toxins incl. Trop. Dis.
Meira,DA; Almeida,RAMB; Barbosa,AN; de Souza,LR; Olivo,TET; Henriques,RMS; Golim,MA; Araújo Jr,JP; Nagoshi,LR; Orikaza,CM; Calvi,SA.
A cross-sectional study was performed on HIV-1 infected individuals with or without antiretroviral treatment (ARV) in the AIDS Day Hospital, Botucatu Medical School, UNESP. Between August 2004 and October 2005, 73 HIV-1 infected individuals were divided into three groups: infected individuals with or without AIDS who had never received ARV (G1 = 15); patients on HAART that had had plasma HIV-1 RNA viral load (VL) equal to or greater than 50 copies/mL (G2 = 27); and patients on HAART with undetectable VL for at least the past six months (G3 = 31). There was also an additional group that comprised blood donors without any sign of the disease and with negative HIV serum tests (G4 = 20), which was the control group. Serum cytokine levels (values in pg/mL) were...
Tipo: Info:eu-repo/semantics/article Palavras-chave: RT-PCR; ELISA; Cytokines; HIV; AIDS; HAART; Apoptosis.
Ano: 2008 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992008000400011
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Avaliação da urina e de leucócitos como amostras biológicas para a detecção ante mortem do vírus da cinomose canina por RT-PCR em cães naturalmente infectados Arq. Bras. Med. Vet. Zootec.
Negrão,F.J.; Alfieri,A.A.; Alfieri,A.F..
Urine and leucocytes were comparatively evaluated as clinical samples for ante mortem detection of the canine distemper virus (CDV) by a reverse transcription-polymerase chain reaction (RT-PCR) assay. One hundred and eighty eight dogs with clinical symptoms of distemper, were distributed in three groups. The group A was constituted of 93 dogs with systemic signs of distemper; the group B by 11 dogs with neurological signs, and the group C by 84 dogs that presented simultaneously systemic and neurological signs. In 66.5% (125/188) of the dogs was amplified an amplicon with 287 base pair of the CDV nucleoprotein gene. In 60.8% (76/125) of the animals the CDV was detected simultaneously in the urine and leucocytes, and in 39.2% (49/125) of the dogs just a...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cão; Cinomose; Diagnóstico; RT-PCR.
Ano: 2007 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352007000100042
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Avaliação de técnicas biomoleculares em ovócitos e amostras de sangue de fêmeas infectadas com vírus da Artrite Encefalite Caprina. Repositório Alice
SILVA, P. A. F. da; SOUSA, S. D.; SOUSA, K. C. de; VERAS, A. K. A.; CAVALCANTE, F. R. A.; ANDRIOLI, A..
Tipo: Parte de livro Palavras-chave: CAE; Vírus da artrite encefalite caprina; CAEV; PCR; RT-PCR; Artrite encefalite caprina; Animal disease.; Ovócito; Caprino; Cabra; Doença animal; Diagnostico.; Goats; Caprine arthritis-encephalitis..
Ano: 2012 URL: http://www.alice.cnptia.embrapa.br/alice/handle/doc/938408
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Avaliação de três kits comerciais para extração de RNA de Influenza A em fluido oral de suínos. Repositório Alice
BARCELLOS, D. E. S. N.; LARA, A. C.; GAVA, D.; SCHAEFER, R.; GOSLAR, M. S..
Tipo: Artigo de periódico Palavras-chave: Detecção; RT-PCR; Suinocultura; Vírus; Influenza A virus.
Ano: 2021 URL: http://www.alice.cnptia.embrapa.br/alice/handle/doc/1137274
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Breakdown of resistance in sweet pepper against Pepper yellow mosaic virus in Brazil Scientia Agricola
Gioria,Ricardo; Braga,Renato de Souza; Krause-Sakate,Renate; Roullier,Caroline; Rosa,Daniel Dias; Moura,Monika Fecury; Souza-Dias,José Alberto Caram de; Sawazaki,Haiko Enok; Camargo,Luis Eduardo Aranha; Rezende,Jorge Alberto Marques.
Plants of Capsicum annuum cv. Magali R, resistant to Pepper yellow mosaic virus (PepYMV), which showed severe yellow mosaic, leaf malformation and stunting were observed during the 2003/04 growing season in Lins, São Paulo State, Brazil. Potyvirus-like particles observed in leaf sap from infected plants under the electron microscope reacted with an antiserum against PepYMV in PTA-ELISA. In addition to C. annuum cv. Magali R, this potyvirus also infected systemically the resistant C. annuum cv. Rubia R. The nucleotide sequence of part of the CP gene of this potyvirus shared 96-98% identity with that of other PepYMV isolates. The partial nucleotide sequence of the 3' NTR showed 94-96% identity with that of PepYMV. These data indicate that this potyvirus is a...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Potyvirus; RT-PCR; ELISA; Resistance gene.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-90162009000200017
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Canine distemper diagnosis by RT-PCR in dog samples from São Paulo State submitted to rabies laboratory diagnosis MV&Z
Macedo, Carla Isabel; Peixoto, Zélia Maria Pinheiro; Castilho, Juliana Galera; Oliveira, Rafael de Novaes; Rodrigues, Adriana Cândido; Achkar, Samira Maria.
Canine distemper (CD) is one of the most important infectious diseases in domestic dogs. In Brazil, CD is still the principal cause of mortality of dogs in some urban populations. Although different viral gene sequences have been identified for detecting the canine distemper virus (CDV), the N gene appears to be a better target for the amplification of specific fragments from all CDV strains. Using RT-PCR targeted to the CDV N gene, during the year 2014 we analyzed 190 central nervous system (CNS) samples of dogs from the state of São Paulo, Brazil, with symptoms suggestive of encephalitis and sent them to the Pasteur Institute for the diagnosis of rabies. Positivity for CD was over 50%, indicating that the disease remains an important cause of canine...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Canine Distemper; RT-PCR; Encephalitis; Diagnosis cinomose canina; RT-PCR; Encefalite; Diagnóstico.
Ano: 2016 URL: http://www.revistamvez-crmvsp.com.br/index.php/recmvz/article/view/31032
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CaV 3.1 and CaV 3.3 account for T-type Ca2+ current in GH3 cells BJMBR
Mudado,M.A.; Rodrigues,A.L.; Prado,V.F.; Beirão,P.S.L.; Cruz,J.S..
T-type Ca2+ channels are important for cell signaling by a variety of cells. We report here the electrophysiological and molecular characteristics of the whole-cell Ca2+ current in GH3 clonal pituitary cells. The current inactivation at 0 mV was described by a single exponential function with a time constant of 18.32 ± 1.87 ms (N = 16). The I-V relationship measured with Ca2+ as a charge carrier was shifted to the left when we applied a conditioning pre-pulse of up to -120 mV, indicating that a low voltage-activated current may be present in GH3 cells. Transient currents were first activated at -50 mV and peaked around -20 mV. The half-maximal voltage activation and the slope factors for the two conditions are -35.02 ± 2.4 and 6.7 ± 0.3 mV (pre-pulse of...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Calcium channel; T-type; Electrophysiology; RT-PCR; GH3 cells.
Ano: 2004 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2004000600020
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Characterization of a new potyvirus causing mosaic and flower variegation in Catharanthus roseus in Brazil Scientia Agricola
Maciel,Sheila Conceição; Silva,Ricardo Ferreira da; Reis,Marcelo Silva; Jadão,Adriana Salomão; Rosa,Daniel Dias; Giampan,José Segundo; Kitajima,Elliot Watanabe; Rezende,Jorge Alberto Marques; Camargo,Luis Eduardo.
Catharanthus roseus is a perennial, evergreen herb in the family Apocynaceae, which is used as ornamental and for popular medicine to treat a wide assortment of human diseases. This paper describes a new potyvirus found causing mosaic symptom, foliar malformation and flower variegation in C. roseus. Of 28 test-plants inoculated mechanically with this potyvirus, only C. roseus and Nicotiana benthamiana developed systemic mosaic, whereas Chenopodium amaranticolor and C. quinoa exhibited chlorotic local lesions. The virus was transmitted by Aphis gossypii and Myzus nicotianae. When the nucleotide sequence of the CP gene (768nt) was compared with other members of the Potyviridae family, the highest identities varied from 67 to 76 %. For the 3' UTR (286nt),...
Tipo: Info:eu-repo/semantics/article Palavras-chave: RT-PCR; Apocynaceae; Periwinkle; Diagnose; Genome sequencing.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-90162011000600013
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Characterization of Citrus tristeza virus isolates from Colombia Rev. Protección Veg.
Morales,Juan; Orlando,Acosta; Tamayo,Pablo; Peñaranda,José.
Isolates of Citrus tristeza virus around the world differ widely in their biological properties. These properties may depend on strain mixtures within individual trees. Monoclonal Antibody 13 (MCA13) is an antibody that detects almost all the severe strains causing Quick Decline and Stem Pitting CTV syndromes. The objectives of this work were: I) to determine the percentage of CTV isolates reactive to the MCA13 antibody in each Colombian region evaluated and II) to analyze the genetic diversity of isolates from these regions. In almost all the regions evaluated, CTV MCA13 reactive isolates were identified; however, only one was found in Mompox area. The coat protein gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and analyzed...
Tipo: Journal article Palavras-chave: Virus cross-protection; Citrus tristeza disease; Citrus spp; RT-PCR; SSCP.
Ano: 2013 URL: http://scielo.sld.cu/scielo.php?script=sci_arttext&pid=S1010-27522013000100006
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Cloprostenol administration in the first week postpartum reduces expression of oxytocin receptors in the endometrium in Holstein-Zebu cows Arq. Bras. Med. Vet. Zootec.
Camargos,A.; Wohlres-Viana,S.; Costa,I.F.; Camargo,L.S.; Ferreira,J.C.; Ramos,A.A.; Oba,E..
ABSTRACT The present study investigated the hormonal profile and expression of prostaglandin F2α (PGF2α), oxytocin and estrogen receptors in uterine tissues of postpartum cows treated with cloprostenol. Twenty Holstein-Zebu crossbred cows were treated with saline solution (treatment CONT) or cloprostenol (treatment CLO), both administered two and five days postpartum. Blood samples were collected on days two, seven, 14, 21 and 28 postpartum for progesterone, PGF2α metabolite (PGFM) and estradiol determination, and endometrial biopsy was performed in order to quantify the expression of oxytocin receptor (OXTR), prostaglandin F receptor (PTGFR) and estrogen receptor 1 (ERS1) genes. In the CLO treatment, expression of OXTR was reduced (P<0.05) but no...
Tipo: Info:eu-repo/semantics/article Palavras-chave: PTGFR; ESR1; Progesterone; Estradiol; RT-PCR.
Ano: 2017 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352017000400821
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Comparação entre ELISA e RT-PCR na detecção de vírus em parreirais no Nordeste do Brasil. Repositório Alice
PIO-RIBEIRO, G.; ANDRADE, G. P.; CATARINO, A. D. M.; FAJARDO, T. V. M.; NICKEL, O.; BARBOSA, M. A. G.; TAVARES, S. C. C. de H.; LEAL, C. M..
Resumo apresentado de número 604-1.
Tipo: Parte de livro Palavras-chave: RT-PCR; Comparação; Detecção; Região Nordeste; Brasil.; Viticultura; Uva; Doença de planta; Vírus; Elisa..
Ano: 2013 URL: http://www.alice.cnptia.embrapa.br/alice/handle/doc/979488
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COMPARACIÓN DEL LÍMITE DE DETECCIÓN DE MÉTODOS DE ANÁLISIS DE ÁCIDOS NUCLEICOS PARA EL VIROIDE DEL ENANISMO DEL LÚPULO (HSVd) EN CÍTRICOS Rev. Protección Veg.
Soto,Marianela; González,Lien; Peralta,Esther Lilia; Duran-Vila,Nuria.
Se determinó y comparó el límite de detección de la electroforesis secuencial en geles de poliacrilamida (sPAGE), hibridación de ácidos nucleicos (NASH) y transcripción reversa-reacción en cadena de la polimerasa (RT-PCR) para el diagnóstico del viroide del enanismo del lúpulo (HSVd) en cítricos, basados en el análisis de ácidos nucleicos de plantas de cidro y pepino como amplificadores biológicos. El límite de detección de la sPAGE fue mayor (5x10-3) en plantas de pepino que en plantas de cidro (5x10-2); sin embargo, este fue 50 veces menos sensible que el de la NASH (10-4). El límite de detección de la RT-PCR en pepino fue el mayor de todos los métodos analizados (10-6). Dicho valor se potenció cuando los productos de RT-PCR fueron detectados por NASH en...
Tipo: Journal article Palavras-chave: Viroide; HSVd; SPAGE; NASH; RT-PCR; Diagnóstico molecular.
Ano: 2007 URL: http://scielo.sld.cu/scielo.php?script=sci_arttext&pid=S1010-27522007000200005
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Comparison of different methods for total RNA extraction from sclerotia of Rhizoctonia solani Electron. J. Biotechnol.
Shu,Canwei; Sun,Si; Chen,Jieling; Chen,Jianyi; Zhou,Erxun.
Background Rhizoctonia solani (teleomorph: Thanatephorus cucumeris) is one of the most important pathogens of rice (Oryza sativa L.) that causes severe yield losses in all rice-growing regions. Sclerotia, formed from the aggregation of hyphae, are important structures in the life cycles of R. solani and contain a large quantity of polysaccharides, lipids, proteins and pigments. In order to extract high-quality total RNA from the sclerotia of R. solani, five methods, including E.Z.N.A.&#8482; Fungal RNA Kit, sodium dodecyl sulfate (SDS)-sodium borate, SDS-polyvinylpyrrolidone (PVP), guanidinium thiocyanate (GTC) and modified Trizol, were compared in this study. Results The electrophoresis results showed that it failed to extract total RNA from the...
Tipo: Journal article Palavras-chave: Polyvinylpyrrolidone; Rhizoctonia solani; RT-PCR; Sclerotia; Sodium dodecyl sulfate; Total RNA extraction.
Ano: 2014 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582014000100009
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Comparison of four extraction methods to detect hepatitis A virus RNA in serum and stool samples BJID
Paula,Vanessa Salete de; Villar,Livia Melo; Gaspar,Ana Maria Coimbra.
The efficiency of extraction methods for hepatitis A virus (HAV) RNA in clinical samples is of great importance for molecular diagnosis, especially in regions endemic for HAV, such as Brazil. We compared the efficiency of four different extraction techniques in serum and stool samples for the detection of hepatitis A virus by reverse transcription PCR (RT-PCR). We used PCR to analyse serum and stool samples of 12 patients who were referred to the Brazilian Reference Center for Viral Hepatitis (BRCVH) in Rio de Janeiro. The methods tested were Proteinase K, Silica, TRIzol and Guanidine isothiocyanate. Proteinase K extraction was the best method for serum samples; it detected the HAV-RNA in 11 of the 12 samples. The guanidine isothiocyanate method was the...
Tipo: Info:eu-repo/semantics/article Palavras-chave: HAV; Extraction methods; Serum and stool; RT-PCR.
Ano: 2003 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702003000200007
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