Corey E, Arfman E W, Liu A Y, Vessella R L
Department of Urology, School of Medicine, University of Washington, Seattle 98195, USA.
Clin Chem. 1997 Mar;43(3):443-52.
The possibility of improving diagnosis of micrometastases from prostate cancer by further enhancing the detection of prostate-specific antigen-producing cells in circulation is being evaluated. We have developed a reverse transcriptase-PCR protocol with the desirable characteristics of low limit of detection, high specificity, reproducibility of response, and ease of performance. Among the procedural alterations that have contributed to these improvements are longer PCR primers, a two-step amplification cycle, and hot-start PCR. We have lowered the limit of detection to one LNCaP prostate-cancer cell in 10(8) peripheral blood mononuclear cells, and samples of blood and bone marrow from healthy donors have yielded no false positives. Because PCR procedures frequently exhibit tube-to-tube variability, we have incorporated a set of internal and external controls into the protocol-a significant advance in assuring assay reliability.
目前正在评估通过进一步加强对循环中产生前列腺特异性抗原细胞的检测来改善前列腺癌微转移诊断的可能性。我们已经开发出一种逆转录酶聚合酶链反应方案，该方案具有检测下限低、特异性高、反应可重复性好以及操作简便等理想特性。促成这些改进的程序改变包括更长的聚合酶链反应引物、两步扩增循环和热启动聚合酶链反应。我们已将检测下限降低至10⁸个外周血单个核细胞中有1个LNCaP前列腺癌细胞，并且来自健康供体的血液和骨髓样本未产生假阳性结果。由于聚合酶链反应程序经常表现出管间变异性，我们已在方案中纳入一组内部和外部对照——这是确保检测可靠性方面的一项重大进展。
