{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,4,16]],"date-time":"2025-04-16T22:43:35Z","timestamp":1744843415381},"reference-count":19,"publisher":"FapUNIFESP (SciELO)","issue":"5","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Rev. Inst. Med. trop. S. Paulo"],"published-print":{"date-parts":[[2014,9]]},"abstract":"<jats:p>This study describes the development and application of a new PCR assay for the specific detection of pathogenic leptospires and its comparison with a previously reported PCR protocol. New primers were designed for PCR optimization and evaluation in artificially-infected paraffin-embedded tissues. PCR was then applied to post-mortem, paraffin-embedded samples, followed by amplicon sequencing. The PCR was more efficient than the reported protocol, allowing the amplification of expected DNA fragment from the artificially infected samples and from 44% of the post-mortem samples. The sequences of PCR amplicons from different patients showed &gt;99% homology with pathogenic leptospires DNA sequences. The applicability of a highly sensitive and specific tool to screen histological specimens for the detection of pathogenic Leptospira spp. would facilitate a better assessment of the prevalence and epidemiology of leptospirosis, which constitutes a health problem in many countries.<\/jats:p>","DOI":"10.1590\/s0036-46652014000500007","type":"journal-article","created":{"date-parts":[[2014,9,13]],"date-time":"2014-09-13T14:31:50Z","timestamp":1410618710000},"page":"411-415","source":"Crossref","is-referenced-by-count":5,"title":["HIGH SENSITIVE PCR METHOD FOR DETECTION OF PATHOGENIC Leptospira spp. IN PARAFFIN-EMBEDDED TISSUES"],"prefix":"10.1590","volume":"56","author":[{"given":"Angel Alberto","family":"Noda","sequence":"first","affiliation":[{"name":"Institute of Tropical Medicine ?Pedro Kour\u00ed?,  Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Islay","family":"Rodr\u00edguez","sequence":"additional","affiliation":[{"name":"Institute of Tropical Medicine ?Pedro Kour\u00ed?,  Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yaindrys","family":"Rodr\u00edguez","sequence":"additional","affiliation":[{"name":"Institute of Tropical Medicine ?Pedro Kour\u00ed?,  Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anamays","family":"Gov\u00edn","sequence":"additional","affiliation":[{"name":"Institute of Tropical Medicine ?Pedro Kour\u00ed?,  Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carmen","family":"Fern\u00e1ndez","sequence":"additional","affiliation":[{"name":"Institute of Tropical Medicine ?Pedro Kour\u00ed?,  Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ana Margarita","family":"Obreg\u00f3n","sequence":"additional","affiliation":[{"name":"Institute of Tropical Medicine ?Pedro Kour\u00ed?,  Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"530","reference":[{"key":"ref1","first-page":"107","article-title":"Nucleic acid extraction from clinical specimens for PCR applications","volume":"30","author":"Aygan A","year":"2006","journal-title":"Turk J Biol"},{"key":"ref2","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1136\/jcp.2007.046714","article-title":"PCR testing for <em>Treponema pallidum<\/em> in paraffin-embedded skin biopsy specimens: test design and impact on the diagnosis of syphilis","volume":"61","author":"Behrhof W","year":"2008","journal-title":"J Clin Pathol"},{"key":"ref3","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1016\/S0923-2508(03)00166-9","article-title":"Direct detection of leptospiral material in human postmortem samples","volume":"154","author":"Brown PD","year":"2003","journal-title":"Res Microbiol"},{"key":"ref4","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1590\/S0074-02762012000100012","article-title":"Comparison of two PCR methods for detection of <em>Leptospira interrogans<\/em> in formalin-fixed and paraffin-embedded tissues","volume":"107","author":"D'Andrea A","year":"2012","journal-title":"Mem Inst Oswaldo Cruz"},{"key":"ref5","first-page":"171","article-title":"DNA extraction from paraffin embedded tissues by Chelex-100","volume":"39","author":"De Armas Y","year":"2006","journal-title":"Rev Esp Patol"},{"key":"ref6","doi-asserted-by":"crossref","first-page":"1413","DOI":"10.2217\/fmb.10.102","article-title":"<em>Leptospira<\/em> as an emerging pathogen: a review of its biology, pathogenesis and host immune responses","volume":"5","author":"Evangelista KV","year":"2010","journal-title":"Future Microbiol"},{"key":"ref7","doi-asserted-by":"crossref","first-page":"1699","DOI":"10.1111\/j.1365-3156.2006.01727.x","article-title":"Polymerase chain reaction in comparison with serological tests for early diagnosis of human leptospirosis","volume":"11","author":"Fonseca CA","year":"2006","journal-title":"Trop Med Int Health"},{"key":"ref8","doi-asserted-by":"crossref","first-page":"505","DOI":"10.3201\/eid1403.071064","article-title":"Leptospirosis associated Severe Pulmonary Hemorrhagic Syndrome, Salvador, Brazil","volume":"14","author":"Gouveia EL","year":"2008","journal-title":"Emerg Infect Dis"},{"key":"ref9","first-page":"44","article-title":"Leptospiral pneumonia","volume":"12","author":"Hill MK","year":"1997","journal-title":"Semin Respir Infect"},{"issue":"(9181)","key":"ref10","doi-asserted-by":"crossref","first-page":"820","DOI":"10.1016\/S0140-6736(99)80012-9","article-title":"Urban epidemic of severe leptospirosis in Brazil. 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