A novel mutant-enriched liquidchip technology for the qualitative detection of somatic mutations in KRAS gene from both serum and tissue samples

摘要: Background: Somatic mutations in the KRAS gene have been reported to confer drug resistance epidermal growth factor receptor tyrosine kinase inhibitors and some monoclonal antibodies. However, current DNA mutation detection technologies are primarily sequencing-based not high throughput, nor sensitive enough meet clinical needs. Methods: A mutant-enriched PCR method was designed by introducing a unique restriction enzyme site product. This allowed wild-type sequence be selectively removed digestion before application Luminex liquidchip system. Results: total of 100 copies mutant fragment mixed with 1 × 10 5 could detected achieve sensitivity 0.1 %. technology is currently used for testing somatic purpose pharmacogenomic evaluation. Serum samples from 109 patients non-small cell lung cancer were tested 34 (34/109). The formalin-fixed paraffin-embedded 60 colorectal 19 (19/60). Conclusions: novel, qualitative, sensitive, reliable throughput has developed detecting using serum samples.