Comparison of the RNA-amplification based methods RT-PCR and NASBA for the detection of circulating tumour cells.
Comparison of the RNA-amplification based methods RT-PCR and NASBA for the detection of circulating tumour cells.
复制标题
DOI：
10.1038/sj.bjc.6600014
复制
发表时间：
2002-01-07
影响因子：
8.8
通讯作者：
Selby, P
中科院分区：
文献类型：
--
作者：
Burchill, S A;Perebolte, L;Johnston, C;Top, B;Selby, P
逆转录聚合酶链反应（RT-PCR）越来越多地用于检测血液或骨髓中临床上重要的肿瘤细胞。这可能导致重新定义无病和临床复发。然而，其临床应用可能会受到缺乏自动化或可重复性的限制。最近的研究表明，基于核酸序列的靶RNA扩增可能更稳健。在这项研究中，建立了基于核酸序列的扩增检测黑色素瘤，结直肠癌和前列腺癌细胞。基于核酸序列的扩增和RT-PCR均成功扩增了黑色素瘤和结直肠癌患者外周血样本中的目标RNA，但只有RT-PCR检测到前列腺癌患者血液样本中的PSA。通过RT-PCR分析的样品重复之间存在相对较好的一致性（酪氨酸酶的Kappa值为1，CK-20的Kappa值为0.67，PSA的Kappa值为1），但通过基于核酸序列的扩增分析时的一致性较低。这可能会限制NASBA用于检测临床显著疾病的常规使用。总之，RT-PCR似乎是目前最可靠的和可重复的方法，用于检测癌症患者的低水平疾病，虽然前瞻性研究是必要的，以评估不同的分子诊断方法的临床效用。英国癌症杂志（2002）86，102-109。DOI：10.1038/sj/bjc/6600014 www.bjcancer.com © 2002癌症研究运动
Increasingly, reverse transcriptase polymerase chain reaction (RT–PCR) is used to detect clinically significant tumour cells in blood or bone marrow. This may result in a redefinition of disease-free and clinical relapse. However, its clinical utility may be limited by lack of automation or reproducibility. Recent studies have suggested nucleic acid sequence-based amplification of target RNA may be more robust. In this study, nucleic acid sequence-based amplification was established to detect melanoma, colorectal and prostate cancer cells. Nucleic acid sequence-based amplification and RT–PCR both successfully amplified target RNA in peripheral blood samples from patients with melanoma and colorectal cancer, but only RT–PCR detected PSA in blood samples from patients with prostate cancer. There was relatively good agreement between sample replicates analyzed by RT–PCR (Kappa values of one for tyrosinase, 0.67 for CK-20 and one for PSA), but less agreement when analyzed by nucleic acid sequence-based amplification. This may limit the routine use of NASBA for the detection of clinically significant disease. In summary, RT–PCR appears at present to be the most reliable and reproducible method for the detection of low-level disease in cancer patients, although prospective studies are warranted to assess the clinical utility of different molecular diagnostic methods. British Journal of Cancer (2002) 86, 102–109. DOI: 10.1038/sj/bjc/6600014 www.bjcancer.com © 2002 The Cancer Research Campaign