1 resultado para school development project
em Instituto Nacional de Saúde de Portugal
Filtro por publicador
- Aberdeen University (3)
- Abertay Research Collections - Abertay University’s repository (1)
- Academic Archive On-line (Jönköping University; Sweden) (1)
- Academic Archive On-line (Stockholm University; Sweden) (1)
- Academic Research Repository at Institute of Developing Economies (1)
- Adam Mickiewicz University Repository (1)
- AMS Tesi di Dottorato - Alm@DL - Università di Bologna (1)
- AMS Tesi di Laurea - Alm@DL - Università di Bologna (1)
- Applied Math and Science Education Repository - Washington - USA (2)
- Aquatic Commons (20)
- Archimer: Archive de l'Institut francais de recherche pour l'exploitation de la mer (1)
- Archive of European Integration (5)
- Archivo Digital para la Docencia y la Investigación - Repositorio Institucional de la Universidad del País Vasco (2)
- Aston University Research Archive (4)
- Biblioteca Digital da Produção Intelectual da Universidade de São Paulo (1)
- Biblioteca Digital de Teses e Dissertações Eletrônicas da UERJ (5)
- Bioline International (1)
- BORIS: Bern Open Repository and Information System - Berna - Suiça (5)
- Boston College Law School, Boston College (BC), United States (1)
- Brock University, Canada (9)
- Bucknell University Digital Commons - Pensilvania - USA (3)
- Bulgarian Digital Mathematics Library at IMI-BAS (9)
- Cambridge University Engineering Department Publications Database (7)
- CentAUR: Central Archive University of Reading - UK (20)
- Chinese Academy of Sciences Institutional Repositories Grid Portal (4)
- Cochin University of Science & Technology (CUSAT), India (4)
- Coffee Science - Universidade Federal de Lavras (2)
- Comissão Econômica para a América Latina e o Caribe (CEPAL) (5)
- CORA - Cork Open Research Archive - University College Cork - Ireland (2)
- Corvinus Research Archive - The institutional repository for the Corvinus University of Budapest (2)
- CUNY Academic Works (4)
- Dalarna University College Electronic Archive (16)
- Deakin Research Online - Australia (101)
- DI-fusion - The institutional repository of Université Libre de Bruxelles (1)
- Digital Commons - Michigan Tech (1)
- Digital Commons @ DU | University of Denver Research (1)
- Digital Commons at Florida International University (11)
- Digital Peer Publishing (1)
- Digital Repository at Iowa State University (3)
- DigitalCommons@The Texas Medical Center (5)
- Doria (National Library of Finland DSpace Services) - National Library of Finland, Finland (4)
- DRUM (Digital Repository at the University of Maryland) (3)
- Duke University (1)
- eResearch Archive - Queensland Department of Agriculture; Fisheries and Forestry (4)
- Fachlicher Dokumentenserver Paedagogik/Erziehungswissenschaften (13)
- Helda - Digital Repository of University of Helsinki (25)
- Instituto Nacional de Saúde de Portugal (1)
- Instituto Politécnico de Castelo Branco - Portugal (1)
- Instituto Politécnico do Porto, Portugal (5)
- Memoria Académica - FaHCE, UNLP - Argentina (13)
- Memorial University Research Repository (1)
- Ministerio de Cultura, Spain (2)
- Open University Netherlands (3)
- Portal de Revistas Científicas Complutenses - Espanha (4)
- QSpace: Queen's University - Canada (2)
- QUB Research Portal - Research Directory and Institutional Repository for Queen's University Belfast (28)
- Queensland University of Technology - ePrints Archive (429)
- RCAAP - Repositório Científico de Acesso Aberto de Portugal (1)
- RDBU - Repositório Digital da Biblioteca da Unisinos (3)
- ReCiL - Repositório Científico Lusófona - Grupo Lusófona, Portugal (5)
- Repositório Aberto da Universidade Aberta de Portugal (1)
- Repositorio Académico de la Universidad Nacional de Costa Rica (1)
- Repositório Científico da Universidade de Évora - Portugal (2)
- Repositório Científico do Instituto Politécnico de Lisboa - Portugal (1)
- Repositório Científico do Instituto Politécnico de Santarém - Portugal (1)
- Repositorio de la Universidad de Cuenca (1)
- Repositório digital da Fundação Getúlio Vargas - FGV (4)
- Repositório Institucional da Universidade de Aveiro - Portugal (1)
- Repositório Institucional da Universidade Federal do Rio Grande do Norte (1)
- Repositório Institucional UNESP - Universidade Estadual Paulista "Julio de Mesquita Filho" (22)
- Research Open Access Repository of the University of East London. (1)
- RUN (Repositório da Universidade Nova de Lisboa) - FCT (Faculdade de Cienecias e Technologia), Universidade Nova de Lisboa (UNL), Portugal (4)
- SAPIENTIA - Universidade do Algarve - Portugal (2)
- School of Medicine, Washington University, United States (1)
- Universidad de Alicante (3)
- Universidad del Rosario, Colombia (3)
- Universidad Politécnica de Madrid (11)
- Universidade de Lisboa - Repositório Aberto (6)
- Universidade Federal de Uberlândia (2)
- Universidade Federal do Pará (12)
- Universidade Federal do Rio Grande do Norte (UFRN) (5)
- Universidade Metodista de São Paulo (4)
- Universidade Técnica de Lisboa (1)
- Universitat de Girona, Spain (2)
- Universitätsbibliothek Kassel, Universität Kassel, Germany (3)
- Université de Montréal, Canada (5)
- University of Connecticut - USA (1)
- University of Michigan (16)
- University of Queensland eSpace - Australia (6)
- University of Southampton, United Kingdom (2)
- University of Washington (1)
- USA Library of Congress (1)
- WestminsterResearch - UK (3)
- Worcester Research and Publications - Worcester Research and Publications - UK (4)
Resumo:
Diagnostic techniques based on PCR have two major problems: false-positive reactions due to contamination with DNA fragments from previous PCRs (amplicons) and false-negative reactions caused by inhibitors that interfere with the PCR. We have improved our previously reported PCR based on the amplification of a fragment of the Mycobacterium tuberculosis complex-specific insertion element IS6110 with respect to both problems. False-positive reactions caused by amplicon contamination were prevented by the use of uracil-N-glycosylase and dUTP instead of dTTP. We selected a new set of primers outside the region spanned by the formerly used primers to avoid false-positive reactions caused by dTTP-containing amplicons still present in the laboratory. With this new primer set, 16 copies of the IS6110 insertion element, the equivalent of two bacteria, could be amplified 10(10) times in 40 cycles, resulting in a mean efficiency of 77% per cycle. To detect the presence of inhibitors of the Taq polymerase, which may cause false-negative reactions, part of each sample was spiked with M. tuberculosis DNA. The DNA purification method using guanidinium thiocyanate and diatoms effectively removed most or all inhibitors of the PCR. However, this was not suitable for blood samples, for which we developed a proteinase K treatment followed by phenol-chloroform extraction. This method permitted detection of 20 M. tuberculosis bacteria per ml of whole blood. Various laboratory procedures were introduced to reduce failure or inhibition of PCR and avoid DNA cross contamination. We have tested 218 different clinical specimens obtained from patients suspected of having tuberculosis. The samples included sputum (n=145), tissue biopsy samples (n=25), cerebrospinal fluid (n=15), blood (n=14), pleural fluid (n=9), feces, (n=7), fluid from fistulae (n=2), and pus from a wound (n=1). The results obtained by PCR were consistent with those obtained with culture, which is the "gold standard." We demonstrate that PCR is a useful technique for the rapid diagnosis of tuberculosis at various sites.