Rapid and Sensitive Closed-Tube Quantification of Human Interferon-γ mRNA by Reverse Transcription-PCR Utilizing Energy-Transfer Labeled Primers
| العنوان: | Rapid and Sensitive Closed-Tube Quantification of Human Interferon-γ mRNA by Reverse Transcription-PCR Utilizing Energy-Transfer Labeled Primers |
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| المؤلفون: | Dan St. Louis, Glenn Nardone, Hiroshi Uehara, Sandra K. Randall, Yuri Khripin |
| المصدر: | Clinical Chemistry. 46:1500-1501 |
| بيانات النشر: | Oxford University Press (OUP), 2000. |
| سنة النشر: | 2000 |
| مصطلحات موضوعية: | Messenger RNA, Fluorophore, Transition (genetics), Biochemistry (medical), Clinical Biochemistry, Biology, Amplicon, Molecular biology, Fluorescence, law.invention, Reverse transcription polymerase chain reaction, chemistry.chemical_compound, chemistry, law, Primer (molecular biology), Polymerase chain reaction |
| الوصف: | Quantification of cytokine gene expression is essential for understanding the biological roles of cytokines in immune responses. Quantitative reverse transcription-PCR has become popular for the sensitive detection and quantification of mRNA (1). However, the necessity of post-PCR sample processing with the potential for carryover contamination of the amplicon makes it a difficult methodology for routine clinical testing. Using AmplifluorTM technology, we have developed a rapid and extremely sensitive assay for cytokine mRNA quantification. The high signal-to-noise ratio of the assay that is achieved by the unique structure of Amplifluor primers allows for closed-tube fluorescence detection and quantification during PCR amplification (real time) or at endpoint. Elimination of laborious post-PCR sample processing enables high-throughput analysis and greatly reduces the risk of carryover contamination. The Amplifluor primer system is a molecular switch for detecting DNA amplification by energy transfer between fluorophore and quencher (2)(3). The OFF-to-ON transition occurs when the conformation of the Amplifluor primer changes from “closed” intramolecular stem-loop structure to an “open” extended structure. This structural change is achieved when the Amplifluor primers … |
| تدمد: | 1530-8561 0009-9147 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_________::672c9782819f76ceef17ee7d63cd97f6Test https://doi.org/10.1093/clinchem/46.9.1500Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi...........672c9782819f76ceef17ee7d63cd97f6 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15308561 00099147 |
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