Suárez Ana, Fra Joaquín, Alonso Rebeca, Sánchez Rosa, Lacave Angel J, Gutiérrez Carmen
Department of Functional Biology, Hospital Central de Asturias, Universidad de Oviedo, Spain.
Anticancer Res. 2002 Mar-Apr;22(2B):1091-5.
Detection of melanoma cells in the peripheral blood of melanoma patients by reverse transcription polymerase chain reaction techniques has demonstrated varying detection rates. This study examined the sensitivity of the technique by employing a modification of the currently used protocols to detect mRNA markers. RT-PCR of tyrosinase and MART-1 was performed after poly A+-RNA isolation from unmanipulated whole blood lysed in the presence of nuclease inhibitors. We found a preclinical sensitivity of 1 GR-M melanoma cell spiked in 1 ml of blood. The clinical sensitivity was tested by studying 22 melanoma patients with advanced disease. The rate of positivity in all patients was 63.6% for tyrosinase, 50% for MRT-1 and 77.3% for at least one molecular marker. This figure increased to 89.5% when considering only those patients with evident macroscopic disease. We can conclude that the technical modifications introduced in this protocol significantly increased the clinical sensitivity compared with other published methods.
通过逆转录聚合酶链反应技术检测黑色素瘤患者外周血中的黑色素瘤细胞，结果显示检测率各不相同。本研究通过对当前使用的方案进行改进以检测mRNA标志物，从而检验该技术的敏感性。在存在核酸酶抑制剂的情况下裂解未经处理的全血后，分离出聚腺苷酸加尾RNA，然后进行酪氨酸酶和黑色素瘤抗原-1（MART-1）的逆转录聚合酶链反应（RT-PCR）。我们发现，在1毫升血液中加入1个GR-M黑色素瘤细胞时该技术的临床前敏感性。通过研究22例晚期黑色素瘤患者来测试临床敏感性。所有患者中，酪氨酸酶的阳性率为63.6%，MRT-1为50%，至少一种分子标志物为77.3%。仅考虑那些有明显肉眼可见病变的患者时，这一数字增至89.5%。我们可以得出结论，与其他已发表的方法相比，本方案中引入的技术改进显著提高了临床敏感性。
