{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T22:15:23Z","timestamp":1792880123540,"version":"4.5.0"},"reference-count":41,"publisher":"American Society for Microbiology","issue":"1","license":[{"start":{"date-parts":[[2003,1,1]],"date-time":"2003-01-01T00:00:00Z","timestamp":1041379200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Clin Microbiol"],"published-print":{"date-parts":[[2003,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            In order to overcome some of the limitations of conventional microbiological techniques in the diagnosis of human brucellosis, a simple PCR-enzyme-linked immunosorbent assay (PCR-ELISA) was developed. After amplification of a 223-bp sequence of a gene that codes for the synthesis of an immunogenetic membrane protein specific for the\n            <jats:italic>Brucella<\/jats:italic>\n            genus (BCSP31), the digoxigenin-labeled amplified product was hybridized with a biotinylated capture probe which was complementary to the inner part of the amplicon. The hybrid was captured on streptavidin-coated microtiter plates and detected by using an antidigoxigenin Fab-peroxidase conjugate. The detection limit of the PCR-ELISA in a background of 3.5 \u03bcg of human genomic DNA was 10 fg (two bacterial cells). The PCR-ELISA showed an analytical sensitivity higher than that of ethidium bromide staining and equal to that obtained by conventional PCR followed by dot blot hybridization. In 59 peripheral blood samples from 57 consecutive patients with active brucellosis and 113 control samples, the PCR-ELISA was found to be 94.9% sensitive and 96.5% specific, whereas the sensitivity of the blood culture was only 70.1%. Since the assay can be performed in 1 day, is very reproducible, is easily standardized, and avoids the risk of infection in laboratory workers, this PCR-ELISA seems to be a practical and reliable tool for the diagnosis of human brucellosis.\n          <\/jats:p>","DOI":"10.1128\/jcm.41.1.144-148.2003","type":"journal-article","created":{"date-parts":[[2003,1,7]],"date-time":"2003-01-07T18:03:11Z","timestamp":1041962591000},"page":"144-148","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":62,"title":["Development and Evaluation of a PCR-Enzyme-Linked Immunosorbent Assay for Diagnosis of Human Brucellosis"],"prefix":"10.1128","volume":"41","author":[{"given":"Pilar","family":"Morata","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Faculty of Medicine"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mari\u0301a I.","family":"Queipo-Ortun\u0303o","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Faculty of Medicine"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jose M.","family":"Reguera","sequence":"additional","affiliation":[{"name":"Infectious Diseases Unit, Internal Medicine Department"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miguel A.","family":"Garci\u0301a-Ordon\u0303ez","sequence":"additional","affiliation":[{"name":"Infectious Diseases Unit, Internal Medicine Department"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana","family":"Ca\u0301rdenas","sequence":"additional","affiliation":[{"name":"Microbiology Service, Carlos Haya University Hospital, Ma\u0301laga, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan D.","family":"Colmenero","sequence":"additional","affiliation":[{"name":"Infectious Diseases Unit, Internal Medicine Department"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","unstructured":"Alton G. 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