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In our study, 88% of the women had their mutation status confirmed, and we excluded known sporadic cases.
Control samples for pyrosequencing constituted 11 PTC samples with BRAF T1799A mutation status confirmed by Sanger sequencing as previously reported for ten of the cases by Sofiadis et al. (29), as well as three parathyroid adenomas.
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IDH mutation status was confirmed using direct sequencing [ 42].
This 'dose-dependent' effect of smoking on EGFR mutation status is confirmed by our findings.
FLCN mutation status was confirmed through DNA extraction from tumor samples and sequencing, as described previously [ 33], using primer sequences from Nickerson et al. [ 13].
KRAS mutation status was confirmed before randomisation by an AstraZeneca-appointed central laboratory with TheraScreen K-RAS ARMS Mutation Kit (Qiagen (formerly DxS Ltd, Manchesterr, UK), or by an AstraZeneca-agreed local laboratory with agreed methods, or an accredited commercial laboratory with proprietary-accredited methods.
Since patients with a low abundance of EGFR mutations may achieve more clinical benefits from EGFR-TKI treatmore, more sensitive and advanced assays, such as droplet-digital PCR (next-generation quantitative PCR) or next-generation sequencing [ 46, 47], may be a preferable option to more accurately evaluate EGFR mutation status and confirm "true" wild-type EGFR NSCLC.
†Smoking status confirmed via cotinine test.
In addition, BRCA2 mutation status has been confirmed as an independent prognostic factor for poorer outcome [7].
For laboratory evidence, whether or not differences exist in the data endpoints in relation to EGFR mutation status should be confirmed in more than one cell line of each type to make any conclusions.
Large-scale multivariate analyses, using pooled data from prospective phase II or III trials in which the EGFR mutation status was clearly confirmed, are needed to clarify this point.
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