{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,11]],"date-time":"2026-02-11T15:07:09Z","timestamp":1770822429656,"version":"3.50.1"},"reference-count":47,"publisher":"American Society for Microbiology","issue":"12","license":[{"start":{"date-parts":[[2002,12,1]],"date-time":"2002-12-01T00:00:00Z","timestamp":1038700800000},"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":[[2002,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Accurate laboratory tests for the diagnosis of active human herpesvirus 8 (HHV-8) infection are becoming essential to study the pathogenesis of HHV-8-associated tumors and for the clinical management of HHV-8-infected individuals. We have developed a highly sensitive, calibrated quantitative real-time PCR assay for the measurement of cell-free HHV-8 DNA in body fluids, based on the addition of a synthetic DNA calibrator prior to DNA extraction. The calibrator controls each sample for the presence of PCR inhibitors, determines a cutoff value of sensitivity for negative samples, and normalizes positive samples for the efficiency of DNA recovery. The assay shows a wide dynamic range of detection (between 1 and 10\n            <jats:sup>6<\/jats:sup>\n            viral genome equivalents\/reaction) and a high degree of accuracy even in the presence of high amounts (up to 1 \u03bcg) of human genomic DNA. Moreover, the assay has a very high sensitivity (lower detection limit, 10 genome equivalents\/ml) and a high degree of reproducibility and repeatability with a coefficient of variation (CV) of &lt;15 and 23%, respectively. Furthermore, the use of the calibrator improves the accuracy of quantitation and decreases the intersample variability (CV, 9 and 6%, respectively). The sensitivity and specificity of the assay were tested with a series of clinical specimens obtained from patients affected by various HHV-8-related diseases, as well as from a wide number of controls. In conclusion, our calibrated real-time PCR assay provides a reliable high-throughput method for quantitation of HHV-8 DNA in clinical and laboratory specimens.\n          <\/jats:p>","DOI":"10.1128\/jcm.40.12.4652-4658.2002","type":"journal-article","created":{"date-parts":[[2002,11,26]],"date-time":"2002-11-26T22:43:39Z","timestamp":1038350619000},"page":"4652-4658","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":40,"title":["Calibrated Real-Time PCR Assay for Quantitation of Human Herpesvirus 8 DNA in Biological Fluids"],"prefix":"10.1128","volume":"40","author":[{"given":"Francesco","family":"Broccolo","sequence":"first","affiliation":[{"name":"Unit of Human Virology, DIBIT, San Raffele Scientific Institute, 20132 Milan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giuseppe","family":"Locatelli","sequence":"additional","affiliation":[{"name":"Unit of Human Virology, DIBIT, San Raffele Scientific Institute, 20132 Milan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Loredana","family":"Sarmati","sequence":"additional","affiliation":[{"name":"Department of Public Health and Cellular Biology, University \u201cTor Vergata,\u201d 00133 Rome"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sara","family":"Piergiovanni","sequence":"additional","affiliation":[{"name":"Unit of Human Virology, DIBIT, San Raffele Scientific Institute, 20132 Milan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fabrizio","family":"Veglia","sequence":"additional","affiliation":[{"name":"I.S.I. Foundation Villa Gualino, 10133 Turin"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Massimo","family":"Andreoni","sequence":"additional","affiliation":[{"name":"Department of Public Health and Cellular Biology, University \u201cTor Vergata,\u201d 00133 Rome"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stefano","family":"Butto\u0300","sequence":"additional","affiliation":[{"name":"Laboratory of Virology, Istituto Superiore di Sanita\u0300, 00161 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Barbara","family":"Ensoli","sequence":"additional","affiliation":[{"name":"Laboratory of Virology, Istituto Superiore di Sanita\u0300, 00161 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paolo","family":"Lusso","sequence":"additional","affiliation":[{"name":"Unit of Human Virology, DIBIT, San Raffele Scientific Institute, 20132 Milan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mauro S.","family":"Malnati","sequence":"additional","affiliation":[{"name":"Unit of Human Virology, DIBIT, San Raffele Scientific Institute, 20132 Milan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1086\/315456"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1097\/00002030-200009080-00004"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.43.2.377"},{"key":"e_1_3_2_5_2","first-page":"207","volume":"1","year":"1998","unstructured":"Calabro, M. 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