Applying and testing the conveniently optimized enzyme mismatch cleavage method to clinical DNA diagnosis

摘要: Abstract Establishing a simple and effective mutation screening method is one of the most compelling problems with applying genetic diagnosis to clinical use. Because there no reliable inexpensive system, amplifying by PCR performing direct sequencing every coding exon gold standard strategy even today. However, this approach expensive time consuming, especially when gene size or sample number large. Previously, we developed CEL nuclease mediated heteroduplex incision polyacrylamide gel electrophoresis silver staining (CHIPS) as an ideal system constructed only conventional apparatuses commercially available reagents. In study, evaluated utility CHIPS technology for in practice COL2A1 , WRN RPS6KA3 mutations newly diagnosed patients Stickler syndrome (autosomal dominant inheritance), Werner recessive inheritance) Coffin–Lowry (X-linked respectively. all three genes, detected DNA variations including disease causative within day. Direct exons these genes confirmed 100% sensitivity specificity. We demonstrate high sensitivity, cost performance reliability compatibility inheritance modes. its low technology, ready use potentially disseminate any laboratories world.