Tuyển tập các báo cáo nghiên cứu về y học được đăng trên tạp chí y học General Psychiatry cung cấp cho các bạn kiến thức về ngành y đề tài: Quantitative biomarker analysis of synovial gene expression by real-time PCR. | Arthritis Research Therapy Vol 5 No 6 Boyle et al. Research article Open Access Quantitative biomarker analysis of synovial gene expression by real-time PCR David L Boyle1 Sanna Rosengren1 William Bugbee2 Arthur Kavanaugh1 and Gary S Firestein1 Center for Innovative Therapy Division of Rheumatology Allergy and Immunology UCSD School of Medicine La Jolla California USA 2Department of Orthopedics UCSD School of Medicine La Jolla California USA Correspondence David L Boyle e-mail dboyle@ Received 21 Jan 2003 Revisions requested 22 Apr 2003 Revisions received 5 Aug 2003 Accepted 19 Aug 2003 Published 8 Oct 2003 Arthritis Res Ther 2003 5 R352-R360 DOI ar1004 2003 Boyle et al. licensee BioMed Central Ltd Print ISSN 1478-6354 Online ISSN 1478-6362 . This is an Open Access article verbatim copying and redistribution of this article are permitted in all media for any purpose provided this notice is preserved along with the article s original URL. Abstract Synovial biomarker analysis in rheumatoid arthritis can be used to evaluate drug effect in clinical trials of novel therapeutic agents. Previous studies of synovial gene expression for these studies have mainly relied on histological methods including immunohistochemistry and in situ hybridization. To increase the reliability of mRNA measurements on small synovial tissue samples we developed and validated real time quantitative PCR Q-PCR methods on biopsy specimens. RNA was isolated from synovial tissue and cDNA was prepared. Cell-based standards were prepared from mitogen-stimulated peripheral blood mononuclear cells. Real time PCR was performed using TaqMan chemistry to quantify gene expression relative to the cell-based standard. Application of the cellular standard curve method markedly reduced intra- and inter-assay variability and Keywords arthritis biomarker rheumatoid synovium corrected amplification efficiency errors compared with the C t method. The inter-assay coefficient of variation was less .
