CONTEXT: A polymerase chain reaction-based companion diagnostic (cobas 4800 BRAF V600 Mutation Test) was recently approved by the US Food and Drug Administration to select patients with BRAF-mutant metastatic melanoma for treatment with the BRAF inhibitor vemurafenib. OBJECTIVES: (1) To compare the analytic performance of the cobas test to Sanger sequencing by using screening specimens from phase II and phase III trials of vemurafenib, and (2) to assess the reproducibility of the cobas test at different testing sites. DESIGN: Specimens from 477 patients were used to determine positive and negative percent agreements between the cobas test and Sanger sequencing for detecting V600E (1799T>A) mutations. Specimens were evaluated with a massively parallel pyrosequencing method (454) to resolve discordances between polymerase chain reaction and Sanger results. Reproducibility of the cobas test was assessed at 3 sites by using 3 reagent lots and an 8-member panel of melanoma samples. RESULTS: A valid cobas result was obtained for all eligible patients. Sanger sequencing had a failure rate of 9.2% (44 of 477). For the remaining 433 specimens, positive percent agreement was 96.4% (215 of 223) and negative percent agreement, 80% (168 of 210). Among 42 cobas mutation-positive/Sanger V600E-negative specimens, 17 were V600E positive and 24 were V600K positive by 454. The cobas test detected 70% of V600K mutations. In the reproducibility study, a correct interpretation was made for 100% of wild-type specimens and specimens with greater than 5% mutant alleles; V600E mutations were detected in 90% of specimens with less than 5% mutant alleles. CONCLUSIONS: The cobas test (1) had a lower assay failure rate than that of Sanger, (2) was more sensitive in detecting V600E mutations, (3) detected most V600K mutations, and (4) was highly reproducible.
CONTEXT: A polymerase chain reaction-based companion diagnostic (cobas 4800 BRAF V600 Mutation Test) was recently approved by the US Food and Drug Administration to select patients with BRAF-mutant metastatic melanoma for treatment with the BRAF inhibitor vemurafenib. OBJECTIVES: (1) To compare the analytic performance of the cobas test to Sanger sequencing by using screening specimens from phase II and phase III trials of vemurafenib, and (2) to assess the reproducibility of the cobas test at different testing sites. DESIGN: Specimens from 477 patients were used to determine positive and negative percent agreements between the cobas test and Sanger sequencing for detecting V600E (1799T>A) mutations. Specimens were evaluated with a massively parallel pyrosequencing method (454) to resolve discordances between polymerase chain reaction and Sanger results. Reproducibility of the cobas test was assessed at 3 sites by using 3 reagent lots and an 8-member panel of melanoma samples. RESULTS: A valid cobas result was obtained for all eligible patients. Sanger sequencing had a failure rate of 9.2% (44 of 477). For the remaining 433 specimens, positive percent agreement was 96.4% (215 of 223) and negative percent agreement, 80% (168 of 210). Among 42 cobas mutation-positive/Sanger V600E-negative specimens, 17 were V600E positive and 24 were V600K positive by 454. The cobas test detected 70% of V600K mutations. In the reproducibility study, a correct interpretation was made for 100% of wild-type specimens and specimens with greater than 5% mutant alleles; V600E mutations were detected in 90% of specimens with less than 5% mutant alleles. CONCLUSIONS: The cobas test (1) had a lower assay failure rate than that of Sanger, (2) was more sensitive in detecting V600E mutations, (3) detected most V600K mutations, and (4) was highly reproducible.
Authors: S Martín-Algarra; M T Fernández-Figueras; J A López-Martín; A Santos-Briz; A Arance; M D Lozano; A Berrocal; J J Ríos-Martín; E Espinosa; J L Rodríguez-Peralto Journal: Clin Transl Oncol Date: 2013-10-16 Impact factor: 3.405
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Authors: Gang Zheng; Harrison Tsai; Li-Hui Tseng; Peter Illei; Christopher D Gocke; James R Eshleman; George Netto; Ming-Tseh Lin Journal: Am J Clin Pathol Date: 2016-05 Impact factor: 2.493
Authors: Grant A McArthur; Paul B Chapman; Caroline Robert; James Larkin; John B Haanen; Reinhard Dummer; Antoni Ribas; David Hogg; Omid Hamid; Paolo A Ascierto; Claus Garbe; Alessandro Testori; Michele Maio; Paul Lorigan; Celeste Lebbé; Thomas Jouary; Dirk Schadendorf; Stephen J O'Day; John M Kirkwood; Alexander M Eggermont; Brigitte Dréno; Jeffrey A Sosman; Keith T Flaherty; Ming Yin; Ivor Caro; Suzanne Cheng; Kerstin Trunzer; Axel Hauschild Journal: Lancet Oncol Date: 2014-02-07 Impact factor: 41.316
Authors: David J Panka; Elizabeth Buchbinder; Anita Giobbie-Hurder; Aislyn P Schalck; Laleh Montaser-Kouhsari; Alireza Sepehr; Donald P Lawrence; David F McDermott; Rachel Cohen; Alexander Carlson; Jennifer A Wargo; Ryan Merritt; Virginia J Seery; F Stephen Hodi; Anasuya Gunturi; Dennie Fredrick; Michael B Atkins; A John Iafrate; Keith T Flaherty; James W Mier; Ryan J Sullivan Journal: Mol Cancer Ther Date: 2014-10-15 Impact factor: 6.261
Authors: Jonathan C Dudley; Grzegorz T Gurda; Li-Hui Tseng; Derek A Anderson; Guoli Chen; Janis M Taube; Christopher D Gocke; James R Eshleman; Ming-Tseh Lin Journal: Mol Diagn Ther Date: 2014-08 Impact factor: 4.074