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Molecular systematics of the Phyllachorales (ascomycota, fungi) based on 18S ribosomal DNA sequences BABT
Wanderlei-Silva,Denise; Ramalho Neto,Eduardo; Hanlin,Richard.
In order to evaluate the monophyly of the Phyllachorales from a molecular standpoint and elucidate its phylogenetic relationships with other orders, a segment of the 18S rRNA gene from several representatives of the Phyllachorales, including species of Glomerella, Phyllachora, Coccodiella (=Coccostroma), Sphaerodothis, Ophiodothella, as well as Magnaporthe was sequenced. Maximum Parsimony analysis revealed that the Phyllachorales was a polyphyletic assemblage of taxa. None of the other members of the Phyllachorales, which produced either a clypeus or stroma, clustered with Glomerella. Of the taxa examined, was Coccodiella the closest relative of Phyllachora. Magnaporthe was closely related to the Diaporthales. Our 18S rDNA data highly supported Glomerella...
Tipo: Info:eu-repo/semantics/article Palavras-chave: DNA sequencing; 18S ribosomal DNA; PCR; Phyllachorales.
Ano: 2003 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132003000300002
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Polymerase chain reaction (PCR) identification of terverticillate Penicillium species isolated from agricultural soils in eskişehir province BABT
Demirel,Rasime; Sariozlu,Nalan Yilmaz; İlhan,Semra.
In the present study, nine terverticillate Penicillium isolates (P. griseofulfum, P. puberulum, P. crustosum, P. aurantiogriseum, P. chrysogenum, P. primulinum, P. expansum, P. viridicatum, Eupenicillium egyptiacum) from 56 soil samples were characterized genetically by a PCR method. The DNAs of the strains were isolated using the glass beads and vortexing extraction method and then used for PCR amplification with the internal transcribed spacer 1 (ITS1) and ITS4 universal fungal specific primers. The ITS regions of fungal ribosomal DNA (rDNA) were sequenced through the CEQ 8000 Genetic Analysis System. ITS-5.8S sequences obtained were compared with those deposited in the GenBank Database. The results indicated that the identification of Penicillium...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Penicillium; Identification methods; PCR; ITS.
Ano: 2013 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132013000600013
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Detection of Lsr2 gene of Mycobacterium leprae in nasal mucus BABT
Custodio,Luiz Antonio; Saito,Alexandre; Amarante,Marla Karine; Fujita,Thiago Cezar; Perim,Aparecida de Lourdes; Costa,Ivete Conchon; Felipe,Ionice; Jankevicius,Shiduca Itow.
In the present study, nasal mucus from patients with leprosy were analyzed by PCR using specific primers for Lsr2 gene of Mycobacterium leprae. The presence of Lsr2 gene in the nasal mucus was detected in 25.80% of patients with paucibacillari leprosy, and 23.07% of contacts. Despite the absence of clinical features in the contact individuals, it was possible to detect the presence of Lsr2 gene in the nasal mucus of these individuals. Therefore, PCR detection of M. leprae targeting Lsr2 gene using nasal mucus samples could contribute to early diagnosis of leprosy.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Mycobacterium leprae; Leprosy; Hanseniase; Nasal mucus; PCR.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132012000300007
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High quality DNA from human papillomavirus (HPV) for PCR/RFLPs BABT
Wanderlei-Silva,Denise; Nobre,Mariana; Gonzaga,Rosa Silva; Viana,Luciana Silva; Ramalho Neto,Eduardo.
The analysis of DNA in clinical samples for a secure diagnostic has become indispensable nowadays. Techniques approaching isolation of high molecular weigth DNA of HPV could lead to efficient amplification and early clinical diagnosis of the virus DNA by PCR (polymerase chain reaction). We describe a fast, non-toxical, efficient and cheap method for DNA isolation of human papilloma virus (HPV) from cervical smears using guanidine (DNAzol solution). A 450 bp DNA band correponding to the late region (L1) of the virus genome was detected by PCR, showing that the DNAzol extraction soluction generated a good viral DNA yield. The electrophoretic pattern after digestion with restriction endonucleases (RFLPs/PCR) revealed the predominance of HPV-16 and HPV-33 in...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Human papillomavirus; DNA purification; PCR; RFLP.
Ano: 2005 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132005000100006
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Incidence of group A rotavirus in urban and rural areas of the city of Londrina-Brazil, from 1995 to 1997 BABT
Meneghetti,Ângelo Cesar; Bolognini,Andrea Maria; Lauretti,Flávio; Linhares,Rosa Elisa Carvalho; Santos,Norma; Nozawa,Carlos Mitihiko.
Rotaviruses are common pathogens and the causal agents of acute diarrhea among children and young animals. The involvement of rotavirus in human diarrheal disease among population of urban and rural areas of the city of Londrina, Parana was evaluated. Nine hundred and five fecal specimens from persons with diarrhea were studied, being 686 and 219 from urban and rural areas, respectively. Thirty-eight samples (4,2%) were positive for rotavirus by polyacrylamide gel electrophoresis of viral RNA and latex agglutination test of which 36 were from urban and two from rural areas. Out of the positive specimens, 17 strains were further characterized by RT-PCR typing assay, resulting in 16 strains of G1 genotype while one sample was found to be a mixture of G1 and...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Rotavirus; Diarrhea; PAGE; LA; PCR.
Ano: 2001 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132001000300006
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Detection of Genetic Relationship in the Tree of Life of Some Quail Through Molecular Markers Analyses BABT
Deef,Lamiaa Elsayed Mokhtar.
ABSTRACT Quail is an important and interesting group of galliform birds. The Common quail (Coturnix coturnix); the Japanese quail (Coturnix japonica); the Panda quail (Coturnix japonica); the Dotted white quail (Coturnix japonica) and the Bobwhite quail (Colinus virginianus) were used in this study. PCR-RFLP and SDS-proteins were performed to reveal the genetic characterization and genetic relationship of the studied quails. Analysis of fragments generated by digestion of PCR product with restriction enzyme NlaIII recorded highly polymorphic restriction profiles. There is a wide intraspecific COI, SEMA3E and TLX genes variability among the studied quails. Protein bands varied from10 to 18 between quails with minimum number of bands were in the Dotted...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Birds; PCR; Quail; SDS-PAGE; Galliformes.
Ano: 2017 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132017000100425
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Validation of a PCR Assay for Chlamydophila abortus rRNA gene detection in a murine model BABT
Silva-Zacarias,Francielle Gibson da; Alfieri,Amauri Alcindo; Spohr,Kledir Anderson Hofstaetter; Lima,Bruna Azevedo de Carvalho; Negrão,Fábio Juliano; Lunardi,Michele; Freitas,Julio Cesar de.
Chlamydophila abortus (C. abortus) is associated with reproductive problems in cattle, sheep, and goats. Diagnosis of C. abortus using embryonated chicken eggs or immortalized cell lines has a very low sensitivity. Polymerase chain reaction (PCR) assays have been used to detect C. abortus infection in clinical specimens and organ fragments, such as placenta, fetal organs, vaginal secretions, and semen. The aim of this study was to develop a PCR assay for the amplification of an 856-bp fragment of the rRNA gene of the Chlamydiaceae family. The PCR assay was evaluated using organs from 15 mice experimentally infected with the S26/3 reference strain of C. abortus. The results of the rRNA PCR were compared to the results from another PCR system (Omp2 PCR) that...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Bovine; Ovine; Caprine; Reproductive failures; Chlamydophila abortus; PCR.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132009000700014
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Phenotypic and molecular characterization of Staphylococcus xylosus: technological potential for use in fermented sausage BABT
Fiorentini,Ângela Maria; Sawitzki,Maristela Cortez; Bertol,Teresinha Marisa; Brod,Fábio Cristiano Angonesi; Pelisser,Márcia Regina; Arisi,Ana Carolina Maisonnave; Sant'Anna,Ernani Sebastião.
Micrococcaceae strains are applied to fermented sausage as starter cultures, where several members of this family are naturally found. The aim of the present work was to isolate and characterize Staphylococcus xylosus from artisanal sausages produced in South Region of Brazil. From 89 isolates presenting catalase positive and coagulase negative activities, 25 strains were selected for phenotypic characterization. Nine strains identified as Staphylococcus xylosus by API-STAPH were evaluated for their nitrate reduction capacity, which showed satisfactory growth of the strains in the presence of nitrite and sodium chloride, demonstrating their potential for use as starter cultures in fermented sausage. The strains were also evaluated through genus and...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Staphylococcus xylosus; Ermented sausage; Starter cultures; PCR.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132009000300026
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Quality Control of Biotechnological Inputs DetectingMycoplasma BABT
Netto,Cristiane; Soccol,Vanete Thomaz; Sepulveda,Lya; Garcia,Gabriel Henrique Oliveira; Timenetsky,Jorge.
The aim of this work was to study the Polymerase Chain Reaction (PCR) as a tool of quality control of bovine sera and cellular cultures used in the biotechnological industry. A total of 46 samples of bovine sera derived from two slaughterhouses and 33 samples of BHK21 cells derived from two biotechnological industries were evaluated using the primers GPO-3 (sense) and MGSO (antisense). The PCR technique sensibility analysis showed that 280 bp were amplified for the quantities of 50 ng to 0.006 ng of Micoplasma DNA. The primers specificity was confirmed in the test using Staphylococcus aureus, Escherichia coli, Bacillus subtilisand Candida albicans; except by the positive control, none of the samples showed amplification. The presence of Mycoplasma in...
Tipo: Info:eu-repo/semantics/article Palavras-chave: PCR; Bovine serum; BHK21 cells; Mycoplasma; Cellular culture.
Ano: 2015 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132015000200239
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An Alternative and Rapid Method for the Extraction of Nucleic Acids from Ixodid Ticks by Potassium Acetate Procedure BABT
Rodríguez,Islay; Fraga,Jorge; Noda,Angel Alberto; Mayet,Milagro; Duarte,Yanisia; Echevarria,Eduardo; Fernández,Carmen.
Four variants of the potassium acetate procedure for DNA extraction from ixodid ticks at different stage of their life cycles were evaluated and compared with phenol-chloroform and ammonium hydroxide methods. The most rapid and most efficient variant was validated in the DNA extraction procedure from the engorged ticks collected from bovine, canine as well as from house ticks for the screening of Borrelia burgdorferi sensu lato, Anaplasma spp. and Babesia spp. The ammonium hydroxide procedure was used for non-engorged ticks. All the variants were efficient and allowed obtaining PCR-quality material according to the specific amplification of 16S rRNA gene fragment of the original tick. DNA extracted from the ticks under the study was tested by multiplex PCR...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Ticks; PCR; DNA extraction.
Ano: 2014 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132014000400542
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Real time PCR and importance of housekeepings genes for normalization and quantification of mRNA expression in different tissues BABT
Rebouças,Emanuela de Lima; Costa,José Jackson do Nascimento; Passos,Maria Juliane; Passos,José Renato de Sousa; Hurk,Robert van den; Silva,José Roberto Viana.
The aim of this review was to evaluate the importance of the real-time PCR (qRT-PCR) as a technique for mRNA expression analysis in different tissues. Real-time PCR is widely used for quantification of mRNA levels and is a fundamental tool for basic research, molecular medicine and biotechnology.Genes of references are expressed in a wide variety of tissues and cells with minimal variations in their expression levels, and thus are used to normalize data of mRNA quantification. Software programs, such as geNorm, BestKeeper and NormFinder, have been developed to perform the normalization of data, which help to choose the most stable reference gene. Several genes, such as GAPDH, β-actin, β-tubulin, PGK, UBQ, RPL-19 and 18S rRNA have been suggested as...
Tipo: Info:eu-repo/semantics/article Palavras-chave: PCR; Normalization; Reference genes; Gene expression; MRNA.
Ano: 2013 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132013000100019
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A simplified DNA extraction method for PCR analysis of Camarotella spp. BABT
Vitória,Nadja Santos; Bezerra,José Luiz; Gramacho,Karina Peres.
This work aimed to optimize an efficient and simple protocol for DNA extraction of Camarotella species, an obligate plant pathogen that cause verrucosis or "lixa" on coconut tree and other palms, facilitating the molecular studies of these biotrophic microorganisms. The method proposed enabled a fast, reproducible and reliable DNA extraction from Camarotella species.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Camarotella torrendiella; C. acrocomiae; Verrucosis on coconut; DNA; PCR.
Ano: 2010 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132010000200001
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Sexagem de embriões bovinos fecundados in vitro pela técnica de PCR multiplex Braz. J. Vet. Res. Anim. Sci.
LUZ,Marcelo Rezende; WATANABE,Yeda Fumie; FERRO,Jesus Aparecido; FERRO,Maria Inês T.; MAURO,Sônia Marli Singaretti de; HOSSEPIAN DE LIMA,Vera Fernanda Martins; FRANCESCHINI,Paulo Henrique.
Neste trabalho, a técnica de PCR ("polymerase chain reaction") foi utilizada para a sexagem de 92 embriões bovinos fertilizados in vitro. Os embriões originaram-se de fertilização in vitro de oócitos aspirados de ovários de fêmeas bovinas, provenientes de abatedouros comerciais. Os oócitos foram maturados, fertilizados e cultivados até o estádio de blastocisto. Os embriões foram lavados em solução de PBS, transferidos para tubos de polipropileno contendo água ultrapura, e imediatamente congelados a -196ºC. Os embriões foram descongelados sobre isopor contendo gelo picado e tratados com proteinase K. Para a reação de PCR, utilizaram-se alíquotas de 34 µl de cada tudo, onde foram acrescidos dois pares de primers, seqüência BC1.2 e seqüência satélite 1.715,...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Sexagem; Embrião; Bovino; PCR.
Ano: 2000 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-95962000000600006
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Bovine carcass sexing by PCR method Braz. J. Vet. Res. Anim. Sci.
Curi,Rogério Abdallah; Mota,Lígia Souza Lima Silveira da; Amarante,Mônica Regina Vendrame; Lopes,Catalina Romeiro.
The objective of the present study was to develop a methodology that would permit sexing bovine meat ready for commercialization. A male-specific primer sequence was used, followed by analysis of the amplified product. The method proved to be efficient for sex verification and is of practical utility in the prevention of fraud in beef sale.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Bovine; Carcass; Sexing; PCR.
Ano: 2002 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-95962002000300007
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Survey of Mycoplasma pneumoniae in Iranian children with acute lower respiratory tract infections BJID
Hadi,Nahal; Kashef,Sara; Moazzen,Mohammad; Pour,Michel Shamoon; Rezaei,Nima.
OBJECTIVES: Mycoplasma pneumoniae is an atypical pathogen, which is one of the major causes of lower respiratory tract infections (LRTIs) worldwide. This study was performed to determine the role of M. pneumoniae in acute LRTIs in children, who were referred to main pediatric hospitals in Shiraz, Iran, with the diagnosis of LRTI. Polymerase chain reaction method on a throat-swab specimen was utilized to detect M. pneumoniae. RESULTS: One hundred patients with acute LRTIs were investigated in this study. There were 10 positive PCR for M. pneumoniae (10%), including 6 of 62 hospitalized patients and 4 of 38 outpatients. All patients with LRTIs due to M. pneumoniae had cough. Fever, flu like symptoms, dyspnea, pulmonary rales, wheezing, and conjunctivitis...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Atypical bacteria; Mycoplasma pneumoniae; PCR; Respiratory infection.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702011000200001
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The investigation of parvovirus B19 infection in patients with haematological disorders by using PCR and ELISA techniques BJID
Us,Tercan; Ozune,Leyla; Kasifoglu,Nilgun; Akgun,Yurdanur.
Parvovirus B19 has a marked tropism for erythroid progenitor cells. This may lead to chronic anemia in predisposed individuals. The purpose of the study was to investigate the frequency of parvovirus B19 infections in patients with diagnosis of haematological disorders. In order to determine the diagnostic use of different markers of parvovirus B19 infection, serum specimens obtained from 79 patients with haematological disorders were tested for specific antibodies and viral DNA through the use of ELISA and PCR techniques. Evidence of parvovirus B19 infection was found in 23/79 (29.1%) patients by demonstrating viral DNA and/or specific IgM antibody. B19 infection was established in 3 of 11 patients with chronic myeloid leukemia, in 3 of 11 acute myeloid...
Tipo: Info:eu-repo/semantics/article Palavras-chave: PCR; ELISA; Haematological disorder.
Ano: 2007 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702007000300006
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Rapid detection of Vancomycin-Resistant Enterococci (VRE) in rectal samples from patients admitted to intensive care units BJID
d'Azevedo,Pedro Alves; Santiago,Kelly Aline de Souza; Furtado,Guilherme Henrique Campos; Xavier,Diego Batista; Pignatari,Antonio Carlos Campos; Titze-de-Almeida,Ricardo.
The reduction in time required to identify vancomycin-resistant enterococci (VRE) has gained increased importance during hospital outbreaks. In the present study, we implemented a laboratory protocol to speed up the VRE screening from rectal samples. The protocol combines a medium for selective VRE isolation (VREBAC®, Probac, São Paulo) and a multiplex PCR for detection and identification of vanA and vanB resistance genes. The screening performance was analyzed in 114 specimens collected from four intensive care units. The swabs were collected at two periods: (1) during a VRE outbreak (February 2006, n=83 patients) and (2) at the post-outbreak period, after adoption of infection control measures (June 2006, n=31 patients). Forty-one/83 VRE (49.4%) and...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Enterococcus; VRE; Vancomycin; Diagnostic; PCR.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702009000400010
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Western blot seroindeterminate individuals for Human T-lymphotropic Virus 1/2 (HTLV-1/2) in Fortaleza (Brazil): a serological and molecular diagnostic and epidemiological approach BJID
Santos,Terezinha de Jesus Teixeira; Costa,Carlos Maurício de Castro; Goubau,Patrick; Vandamme,Anne-Mieke; Desmyter,Jan; Van Dooren,Sonia; Mota,Rosa M. S.; Costa,Francine Bovy de Castro; Oliveira,Ana C. S.; Gomes,Vânia Barreto A.F.; Carneiro-Proietti,Anna B.; Bruin,Veralice Meireles Sales de; Sousa,Francisca C. F. de; Oriá,Reinaldo Barreto.
How to handle Western blot (WB) seroindeterminate individuals for Human T-lymphotropic Virus 1/2 (HTLV-1/2) constitutes a challenge for blood banks and fam ilies. We made a cross-sectional study of 191 enzyme linked immunoassay (EIA) reactive individuals from the hematological center (HEMOCE) of Fortaleza (Brazil), examining their serological (WB) and molecular (PCR) diagnosis, and demographic profiles, as well as a possible association of their condition with other infectious pathologies and risk factors. Ethical institutional approval and personal consent were obtained. Out of 191 EIA reactive individuals, 118 were WB seroindeterminate and 73 were seropositive for HTLV-1/2. In the PCR analysis of 41 WB seroindeterminate individuals, 9 (22%) were positive...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Epidemiology; PCR; HTLV-1/2; Seroindeterminate individuals; Hepatitis; Syphilis; Blood transfusion.
Ano: 2003 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702003000300005
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Detection of HCV by PCR in serum and PBMC of patients with hepatitis C after treatment BJID
Cavalheiro,Norma de Paula; Filgueiras,Thelma Cristina; Melo,Carlos Eduardo; Morimitsu,Suzana Rie; Araújo,Evaldo Stanislau Affonso de; Tengan,Fátima Mitiko; Barone,Antonio Alci.
Although hepatitis C is mainly hepatotropic, some studies suggest that hepatitis C virus (HCV) infects peripheral blood mononuclear cells (PBMC), using them as a reservoir, which might contribute to the development of resistance to treatment. Fifty-four hepatitis-C patients, who had been submitted to treatment, were selected. Blood samples were collected on the same day for the detection of HCV RNA in serum and PBMC by PCR, using the Amplicor HCV 2.0 assay (Roche Diagnostics). HCV genotyping was performed using the INNO-LiPA HCV kit (Versant, Bayer Diagnostics). HCV RNA was detected in both serum and PBMC in 35 (64%) patients and no RNA in 16 (29.6%). Disagreement between the serum and PBMC results was observed for three patients (5.6%), with HCV RNA being...
Tipo: Info:eu-repo/semantics/article Palavras-chave: PCR; Hepatitis C; PBMC; Treatment.
Ano: 2007 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702007000500006
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Comparative performances of serologic and molecular assays for detecting human T lymphotropic virus type 1 and type 2 (HTLV-1 and HTLV-2) in patients infected with human immunodeficiency virus type 1 (HIV-1) BJID
Campos,Karoline Rodrigues; Gonçalves,Maria Gisele; Costa,Nadia Aparecida; Caterino-de-Araujo,Adele.
ABSTRACT The present study evaluated several techniques currently available (commercial kits and in-house assays) for diagnosing human T lymphotropic viruses types 1 and 2 in two groups of patients enrolled at HIV/AIDS specialized care services in São Paulo: Group 1 (G1), n = 1608, 1237 male/371 female, median age 44.3 years old, majority using highly active antiretroviral therapy (HAART); G2, n = 1383, 930 male/453 female, median age of 35.6 years old, majority HAART naïve. Enzyme immunoassays [(EIA) Murex and Gold ELISA] were employed for human T lymphotropic viruses types 1 and 2 screening; Western blotting (WB), INNO-LIA (LIA), real-time PCR pol (qPCR), and nested-PCR-RFLP (tax) were used to confirm infection. Samples were considered human T...
Tipo: Info:eu-repo/semantics/article Palavras-chave: HTLV-1/2; HIV-1; Coinfection; Diagnosis; Immunoblotting; PCR; HAART.
Ano: 2017 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702017000300297
Registros recuperados: 425
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