{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,3]],"date-time":"2026-05-03T23:43:47Z","timestamp":1777851827452,"version":"3.51.4"},"reference-count":14,"publisher":"SAGE Publications","issue":"3","license":[{"start":{"date-parts":[[1999,3,1]],"date-time":"1999-03-01T00:00:00Z","timestamp":920246400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J Histochem Cytochem."],"published-print":{"date-parts":[[1999,3]]},"abstract":"<jats:p>We developed an amplification detection system in which a universal energy transfer-labeled primer (UniPrimer) is used in combination with any target-specific primer pair. The target specific primers each have a 5\u2032 tail sequence, which is homologous to the 3\u2032 end of the UniPrimer which, in turn, has a hairpin structure on the 5\u2032 end. The hairpin structure brings the fluorophore and quencher into close proximity when the primer is free in solution, providing efficient quenching. When the primer is incorporated into the PCR product, the hairpin structure is unfolded and a fluorescent signal can be detected. Using hepatitis C and human papillomavirus as model systems, this study demonstrates several advantages in the hot-start in situ PCR technique with the UniPrimer system, including target specific detection of one DNA copy per cell without a separate in situ hybridization step and detection of an RNA target by RT in situ PCR without overnight DNase digestion. The UniPrimer-based in situ PCR allows rapid and simple detection of any DNA or RNA target without concern for the background from DNA repair invariably evident in paraffin-embedded tissue when a labeled nucleotide is used.<\/jats:p>","DOI":"10.1177\/002215549904700301","type":"journal-article","created":{"date-parts":[[2011,4,1]],"date-time":"2011-04-01T03:11:07Z","timestamp":1301627467000},"page":"273-279","source":"Crossref","is-referenced-by-count":38,"title":["In Situ Amplification Using Universal Energy Transfer-labeled Primers"],"prefix":"10.1177","volume":"47","author":[{"given":"G.J.","family":"Nuovo","sequence":"first","affiliation":[{"name":"MGN Medical Research Laboratory, Setauket, New York"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R.J.","family":"Hohman","sequence":"additional","affiliation":[{"name":"Intergen Discovery Products, Gaithersburg, Maryland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"G.A.","family":"Nardone","sequence":"additional","affiliation":[{"name":"Intergen Discovery Products, Gaithersburg, Maryland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"I.A.","family":"Nazarenko","sequence":"additional","affiliation":[{"name":"Intergen Discovery Products, Gaithersburg, Maryland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"179","published-online":{"date-parts":[[1999,3,1]]},"reference":[{"key":"bibr1-002215549904700301","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/25.16.3235"},{"key":"bibr2-002215549904700301","doi-asserted-by":"publisher","DOI":"10.1093\/clinchem\/43.5.752"},{"key":"bibr3-002215549904700301","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.88.5.1711"},{"key":"bibr4-002215549904700301","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.88.16.7276"},{"key":"bibr5-002215549904700301","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/25.10.1999"},{"key":"bibr6-002215549904700301","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/25.12.2516"},{"key":"bibr7-002215549904700301","volume-title":"PCR In Situ Hybridization: Protocols and Applications","author":"Nuovo GJ","year":"1997","edition":"3"},{"key":"bibr8-002215549904700301","doi-asserted-by":"publisher","DOI":"10.1101\/gr.2.4.305"},{"key":"bibr9-002215549904700301","doi-asserted-by":"publisher","DOI":"10.1101\/gr.4.2.89"},{"key":"bibr10-002215549904700301","first-page":"1239","volume":"139","author":"Nuovo GJ","year":"1991","journal-title":"Am J Pathol"},{"key":"bibr11-002215549904700301","doi-asserted-by":"publisher","DOI":"10.1097\/00000478-199307000-00005"},{"key":"bibr12-002215549904700301","first-page":"267","volume":"55","author":"Nuovo GJ","year":"1995","journal-title":"Cancer Res"},{"key":"bibr13-002215549904700301","doi-asserted-by":"publisher","DOI":"10.1016\/0076-6879(95)46015-2"},{"key":"bibr14-002215549904700301","doi-asserted-by":"publisher","DOI":"10.1038\/nbt0396-303"}],"container-title":["Journal of Histochemistry &amp; Cytochemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.sagepub.com\/doi\/pdf\/10.1177\/002215549904700301","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.sagepub.com\/doi\/full-xml\/10.1177\/002215549904700301","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.sagepub.com\/doi\/pdf\/10.1177\/002215549904700301","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T22:26:40Z","timestamp":1777501600000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.sagepub.com\/doi\/10.1177\/002215549904700301"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1999,3]]},"references-count":14,"journal-issue":{"issue":"3","published-print":{"date-parts":[[1999,3]]}},"alternative-id":["10.1177\/002215549904700301"],"URL":"https:\/\/doi.org\/10.1177\/002215549904700301","relation":{},"ISSN":["0022-1554","1551-5044"],"issn-type":[{"value":"0022-1554","type":"print"},{"value":"1551-5044","type":"electronic"}],"subject":[],"published":{"date-parts":[[1999,3]]}}}