https://scholars.lib.ntu.edu.tw/handle/123456789/551239
標題: | An international collaboration to harmonize the quantitative plasma Epstein-Barr virus DNA assay for future biomarker-guided trials in nasopharyngeal carcinoma | 作者: | Le Q.-T. Zhang Q. Cao H. Cheng A.-J. Pinsky B.A. RUEY-LONG HONG Chang J.T. CHUN-WEI WANG Tsao K.-C. Lo Y.M.D. Lee N. Ang K.K. Chan A.T.C. Chan K.C.A. |
公開日期: | 2013 | 卷: | 19 | 期: | 8 | 起(迄)頁: | 2208-2215 | 來源出版物: | Clinical Cancer Research | 摘要: | Purpose: Persistently elevated posttreatment plasma EBV DNA is a robust predictor of relapse in nasopharyngeal carcinoma (NPC). However, assay standardization is necessary for use in biomarker-driven trials. We conducted a study to harmonize the method between four centers with expertise in EBV DNA quantitation. Experimental Design: Plasma samples of 40 patients with NPC were distributed to four centers. DNA was extracted and EBV DNA copy number was determined by real-time quantitative PCR (BamHI-W primer/probe). Centers used the same protocol but generated their own calibrators. A harmonization study was then conducted using the same calibrators and PCR master mix and validated with ten pooled samples. Results: The initial intraclass correlations (ICC) for the first 40 samples between each center and the index center were 0.62 [95% confidence interval (CI): 0.39-0.78], 0.70 (0.50-0.83), and 0.59 (0.35-0.76). The largest variability was the use of different PCR master mixes and calibrators. Standardization improved ICC to 0.83 (0.5-0.95), 0.95 (0.83-0.99) and 0.96 (0.86-0.99), respectively, for ten archival frozen samples. For fresh plasma with spiked-in EBV DNA, correlations were more than 0.99 between the centers. At 5 EBV DNA copies per reaction or above, the coefficient of variance (CV) was less than 10% for the cycle threshold (Ct) among all centers, suggesting this concentration can be reliably used as a cutoff for defining the presence of detectable EBV DNA. Conclusions: Quantitative PCR assays, even when conducted in experienced clinical labs, can yield large variability in plasma EBV DNA copy numbers without harmonization. The use of common calibrators and PCR master mix can help to reduce variability. ?2013 AACR. |
URI: | https://www.scopus.com/inward/record.uri?eid=2-s2.0-84877099043&doi=10.1158%2f1078-0432.CCR-12-3702&partnerID=40&md5=e11fdb956cc593e7df025e4e4f4edb5d https://scholars.lib.ntu.edu.tw/handle/123456789/551239 |
ISSN: | 1078-0432 | DOI: | 10.1158/1078-0432.CCR-12-3702 | SDG/關鍵字: | tumor marker; virus DNA; article; bioassay; blood sampling; cancer prognosis; clinical article; controlled study; disease association; DNA determination; DNA extraction; Epstein Barr virus; gene dosage; human; instrument validation; nasopharynx carcinoma; nonhuman; priority journal; public-private partnership; quantitative analysis; real time polymerase chain reaction; standardization; tumor volume; virus concentration; Calibration; DNA, Viral; Epstein-Barr Virus Infections; Herpesvirus 4, Human; Humans; International Cooperation; Laboratories; Nasopharyngeal Neoplasms; Real-Time Polymerase Chain Reaction; Reference Standards; Reproducibility of Results; Sensitivity and Specificity; Tumor Markers, Biological |
顯示於: | 腫瘤醫學研究所 |
在 IR 系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。