Pyrosequencing as an alternative to single-strand conformation polymorphism analysis for detection of N-ras mutations in human melanoma metastases.

摘要: Background: Mutations in codons 12, 13, and 61 of the N-ras gene are common alterations cutaneous malignant melanoma. We evaluated pyrosequencing, a simple rapid method used mainly for single-nucleotide polymorphism analysis, as possible alternative to single-strand conformation (SSCP) analysis sequencing N-ras. Methods: sensitivity accuracy pyrosequencing identification mutations 61. Nucleotide dispensation orders were created produce distinct pyrogram peak profiles most frequent codon 12 respectively. Results: The detection limits two found melanoma, which code Arg Lys, 30% 15%, To evaluate on clinical samples, we performed parallel 82 melanoma metastases using SSCP pyrosequencing. All detected by confirmed also correctly identified Codon 26 samples (32%), whereas no 13. Four types mutations, (17%), Lys (10%), Leu (4%), His (1%), identified. Conclusion: Pyrosequencing is an attractive mutation tumor because it displays same rapid. © 2002 American Association Clinical Chemistry