Establishment of experimental conditions for the rapid detection of mutations in the von Hippel-Lindau gene by parallel temperature gradient gel electrophoresis.
Renal cell carcinoma (RCC) is the most common adult kidney neoplasm and is present in approximately 3% of all adult malignancies. Mutations of the von Hippel-Lindau (VHL) gene had been found in 55-70% of sporadic RCC and appear to be a critical event in the pathogenesis of clear-cell RCC. Several screening procedures for the detection of mutations in the VHL gene such as single-strand conformation polymorphism analysis, Southern blot and polymerase chain reaction (PCR)-employing methods had been used. Recently, the perpendicular temperature gradient gel electrophoresis (TGGE) was successfully applied to detect genetic alterations in the VHL gene. Here, the experimental conditions for parallel TGGE were established. The results on eleven samples with mutations in different exons demonstrate that parallel TGGE can be used as a simple and rapid method to detect mutations in the VHL gene. Several samples can be run at the same time, making parallel TGGE ideal for the screening of a large number of samples.
Hernández
A
A
Bone Marrow Donor Center with Transplantation Immunology and Stem Cell Bank, Heinrich-Heine-University, Medical Center Düsseldorf, Germany.
Meyer
A
A
Enczmann
J
J
Ackermann
R
R
Wernet
P
P
D000328:Adult; D002292:Carcinoma, Renal Cell / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006801:Humans; D007680:Kidney Neoplasms / Q000235:genetics; D012680:Sensitivity and Specificity; D013696:Temperature; D014407:Tumor Cells, Cultured; D006623:von Hippel-Lindau Disease / Q000235:genetics*