V600E mutation and p. V600K mutation both in a melanoma sample and p. V600R mutation of a colorectal tumor. Staining with the p. V600E specific monoclonal antibody detected all evaluated p. V600E Wortmannin msds mutations. 22 of these p. V600E mutated samples were melanoma and two were colorectal tumors. Colomba et al. described in con trast a IHC Inhibitors,Modulators,Libraries failure rate of 7. 2% in a cohort of 111 cases due to equivocal staining. Furthermore, case 25, showing a double mutation in codon 600 and codon 601 of the BRAF gene was scored negative in IHC. This is in concordance with the study of Skorokhod et al. who could not detect the double mutation with the monoclonal VE1 antibody either. Eight cases with non p. V600E mutation were scored as 1 and therefore negative in the IHC. Like for the cobas 4800 BRAF V600 test this p.

V600E specificity constitutes the major limitation of Inhibitors,Modulators,Libraries the IHC for routine diagnostics as a single test. However, the Inhibitors,Modulators,Libraries IHC was not completely specific for the p. V600E mutation as cross reactivity was observed in one case with a p. V600R Inhibitors,Modulators,Libraries mutation that was scored as 2. This is in contrary to most other studies report ing no cross reactivity with non p. V600E mutations. Only Heinzerling et al. found for one sam ple an immunohistochemical cross reactivity with p. V600K mutation. Therefore, in our study this method is char acterized by 100% sensitivity but only 98% specificity. Long et al. showed a sensitivity of 97% and a spe cificity of 98% in a cohort of 100 samples. One case of our study highlighted the importance of immunohistochemical staining prior to DNA extraction for mutational analysis.

Case 7 was wildtype using Sanger sequencing, HRM, and cobas BRAF V600 test in the first extraction. NGS Inhibitors,Modulators,Libraries showed a p. V600E mutation with a 3% allele frequency being under our defined threshold. Sec tions for IHC were cut after the molecular analysis and results were positive with a score of 2 by a senior path ologist. Tumor content increased only slightly compared to the first H E stained slide. Therefore, a second extraction was performed and analysis was repeated. The second extract showed a p. V600E mutation using Sanger sequencing, HRM, NGS as well as cobas BRAF V600 test. In general, Sanger sequencing needs 2 4 working days to produce a report. In contrast, HRM is time and cost sav ing and a major advantage is the prevention of contamina tions as HRM is a close tube process.

But it only serves as screening method not giving the exact mutational status. Advantages of pyrosequencing are that it is more sensitive than Sanger sequencing and the amount of work is http://www.selleckchem.com/products/pazopanib.html lower compared to Sanger sequencing hence no clean up steps of the PCR products is needed but result interpretation is more prone to errors. The cobas 4800 BRAF V600 test is charac terized by an easy and fast performance with a low amount of work. Costs are medium compared to the other eva luated methods.