BACKGROUND Single base-pair substitution mutations in the gene for coagulation factor VIII, procoagulant component (hemophilia A) (F8) account for approximately 50% of severe cases of hemophilia A (HA), and almost all moderate or mild cases. Because F8 is a large gene, mutation screening using denaturing HPLC or DNA sequencing is time-consuming and expensive. METHODS We evaluated high-resolut...

BACKGROUND Common methods for identification of DNA sequence variants use gel electrophoresis or column separation after PCR. METHODS We developed a method for sequence variant analysis requiring only PCR and amplicon melting analysis. One of the PCR primers was fluorescently labeled. After PCR, the melting transition of the amplicon was monitored by high-resolution melting analysis. Differen...

BACKGROUND Complete gene analysis of the cystic fibrosis transmembrane conductance regulator gene (CFTR) by scanning and/or sequencing is seldom performed because of the cost, time, and labor involved. High-resolution DNA melting analysis is a rapid, closed-tube alternative for gene scanning and genotyping. METHODS The 27 exons of CFTR were amplified in 37 PCR products under identical conditi...

We developed a novel rapid assay to detect the gyrA mutations that cause quinolone resistance in typhoid and paratyphoid fever Salmonella spp. using high-resolution melting (Idaho Technology, Salt Lake City, UT) analysis of polymerase chain reaction amplicons. The presence of gyrA mutations led to small but consistent changes in amplicon melting temperatures that allowed quinolone-resistant iso...

Identifying mutations in the TP53 gene is important for cancer prognosis, predicting response to therapy, and determining genetic risk. We have developed a high-throughput scanning assay with automatic calling to detect TP53 mutations in DNA from fresh frozen (FF) and formalin-fixed paraffin-embedded (FFPE) tissues. The coding region of the TP53 gene (exons 2-11) was PCR-amplified from breast c...