Wenzel M, Enczmann J, Uhrberg M, Hernández A, Wiese U, Ackermann R, Schmitz-Draeger B, Ebert T, Wernet P
Knochenmarkspenderzentrale, Heinrich-Heine-Universität Düsseldorf, Germany.
Electrophoresis. 1997 Jan;18(1):45-51. doi: 10.1002/elps.1150180109.
Renal cell carcinoma is the most common neoplastic disease of the adult kidney and occurs in its sporadic and hereditary form. Approximately 57% of all renal carcinomas of the clear cell type analyzed revealed a mutation in the von Hippel-Lindau disease (VHL) gene. In the present work, temperature gradient gel electrophoresis (TGGE) is presented as a rapid and precise polymerase chain reaction (PCR)-employing methodology for the detection of mutations in the VHL gene. The theoretical efficiency of TGGE to detect mutations in every base pair of the gene was calculated. According to computer analysis, at least 92% of all known mutations in the VHL gene are detectable. This calculated figure appears to be in agreement with the experimental results. Primary difficulties in analyzing exon 1 of the VHL gene were overcome by the employment of psoralen-cross-linked PCR fragments. In addition, TGGE analysis was used in screening for possible mutations in thirteen renal carcinoma samples. With this protocol TGGE is successfully added to the array of methods for the screening of VHL mutations.
肾细胞癌是成人肾脏最常见的肿瘤性疾病，以散发性和遗传性形式出现。在所有分析的透明细胞型肾癌中，约57%显示von Hippel-Lindau病（VHL）基因突变。在本研究中，温度梯度凝胶电泳（TGGE）作为一种快速、精确的采用聚合酶链反应（PCR）的方法被用于检测VHL基因突变。计算了TGGE检测该基因每个碱基对突变的理论效率。根据计算机分析，VHL基因中至少92%的已知突变是可检测的。这个计算结果似乎与实验结果一致。通过使用补骨脂素交联的PCR片段克服了分析VHL基因第1外显子的主要困难。此外，TGGE分析用于筛查13个肾癌样本中可能的突变。通过该方案，TGGE成功地加入到VHL突变筛查方法的阵列中。
