{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,14]],"date-time":"2025-10-14T01:18:29Z","timestamp":1760404709429,"version":"3.37.3"},"reference-count":33,"publisher":"Wiley","license":[{"start":{"date-parts":[[2017,1,1]],"date-time":"2017-01-01T00:00:00Z","timestamp":1483228800000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Tuberculosis Research and Treatment"],"published-print":{"date-parts":[[2017]]},"abstract":"<jats:p>Tuberculous meningitis (TBM) is a severe form of extrapulmonary tuberculosis. The aims of this study were to evaluate in-house molecular diagnostic protocols of DNA extraction directly from CSF samples and the targets amplified by qPCR as an accurate and fast diagnosis of TBM. One hundred CSF samples from 68 patients suspected of TBM were studied. Four DNA extraction techniques (phenol-chloroform-thiocyanate guanidine, silica thiocyanate guanidine, resin, and resin with ethanol) were compared and CSF samples were used to determine the best target (<jats:italic>IS6110<\/jats:italic>,<jats:italic>MPB64,<\/jats:italic>and<jats:italic>hsp65\u2009KDa<\/jats:italic>) by qPCR. The extraction protocol using the phenol-chloroform-thiocyanate guanidine showed the best results in terms of quantification and sensitivity of PCR amplification, presenting up to 10 times more DNA than the second best protocol, the silica guanidine thiocyanate. The target that showed the best result for TBM diagnosis was the<jats:italic>IS6110<\/jats:italic>. This target showed 91% sensitivity and 97% specificity when we analyzed the results by sample and showed 100% sensitivity and 98% specificity when we analyzed the results by patient. The DNA extraction with phenol-chloroform-thiocyanate guanidine followed by<jats:italic>IS6110 target<\/jats:italic>amplification has been shown to be suitable for diagnosis of TBM in our clinical setting.<\/jats:p>","DOI":"10.1155\/2017\/5089046","type":"journal-article","created":{"date-parts":[[2017,5,30]],"date-time":"2017-05-30T21:03:52Z","timestamp":1496178232000},"page":"1-6","source":"Crossref","is-referenced-by-count":3,"title":["Comparison of DNA Extraction Protocols and Molecular Targets to Diagnose Tuberculous Meningitis"],"prefix":"10.1155","volume":"2017","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5857-1493","authenticated-orcid":true,"given":"Flavia Silva","family":"Palomo","sequence":"first","affiliation":[{"name":"Special Clinical Microbiology Laboratory (LEMC), Federal University of S\u00e3o Paulo (UNIFESP), S\u00e3o Paulo, SP, Brazil"},{"name":"Central Laboratory of S\u00e3o Paulo Hospital, Federal University of S\u00e3o Paulo (UNIFESP), S\u00e3o Paulo, SP, 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