Virchows Archiv

, Volume 474, Issue 1, pp 71–78 | Cite as

BRAF mutation testing in melanoma: results from a German observational multicenter study

  • Arndt Hartmann
  • Peter Schirmacher
  • William Sterlacci
  • Winfried Koch
  • David B. Liesenfeld
  • Birgit Schif
  • Claus Garbe
Original Article


Quality control of BRAF mutation testing methods used in routine practice is crucial for optimal treatment selection. In this prospective study, we assessed the impact of patient/sample characteristics on BRAF mutation testing results in patients with melanoma, during clinical practice. Data were collected on routine testing practices and documented mutation status in patients with melanoma stages IIIB, IIIC, or IV across 28 diagnostic pathology centers in Germany. Patient/sample data collected included: patient age, location of primary melanoma and metastases, origin of sample, melanoma subtype, and quality of tissue. Statistical influence of patient/sample characteristics on BRAF mutation rate was assessed using multiple logistic regression analyses and statistical models developed to predict the probability of BRAF mutations for individual patient cohorts. Data/samples from 642 patients with melanoma were analyzed. BRAF mutations were documented in 241/642 patients (37.5%). The primary statistical model to predict BRAF mutation rates included: age (continuous), origin of sample, method of mutation analysis, and quality of tissue. Analyses of post hoc collected data identified major deviations between documented mutation rates included in this study vs. routinely recorded mutation rates for three centers. When samples from these centers were excluded, the influence of testing method was no longer statistically significant. The final model included patient age, origin of sample (including metastasis location), and quality of tissue. Once validated in an independent population, this type of model could allow pathology centers to compare the performance of their testing methods with what would be expected based on patient, tumor, and sample characteristics.


BRAF mutation Melanoma Quality control Mutational analysis Multiple logistic regression 



We would like to thank the patients, their families, the nurses, and the investigators who participated in this study (see Online Resource 1 for the list of investigators). Funding for this study was provided by Roche Pharma AG, Germany. Support for third-party writing assistance for this manuscript, furnished by Rachel Johnson, PhD, of Health Interactions, was funded by Roche Pharma AG, Germany.

Author contribution statement

A Hartmann contributed to study design and study protocol development; A Hartmann, P Schirmacher, W Sterlacci, and C Garbe contributed to data acquisition; P Schirmacher, W Sterlacci, W Koch, DB Liesenfeld, and B Schif were involved in data analysis and A Hartmann, P Schirmacher, W Sterlacci, W Koch, DB Liesenfeld, B Schif, and C Garbe in data interpretation; W Koch, DB Liesenfeld, and B Schif were responsible for figure development. All authors were involved in writing the paper and had final approval of the submitted and published versions.


This study was funded by Roche Pharma AG, Grenzach-Wyhlen, Germany.

Compliance with ethical standards

Conflict of interest

A Hartmann has received research funding from Sysmex, BioNTech, Nanostring, and Novartis, and has received honoraria from Roche, AstraZeneca, BMS, and MSD, and payment for consultancy or advisory roles from Medoc, Roche, AstraZeneca, BMS, and MSD. P Schirmacher has received research funding from Roche, AstraZeneca, Novartis, Chugai, Thermo Fisher, and Sanofi Aventis, honoraria from Novartis, AstraZeneca, and Roche, and payment for consultancy or advisory roles from Roche, AstraZeneca, Novartis, BMS, MSD, Merck, and Amgen. W Sterlacci has received payment for consultancy or advisory roles from Roche. W Koch is an employee of BDS Koch, which receives payment for statistical services from Roche. B Schif and DB Liesenfeld are employed by Roche. C Garbe has received research funding from BMS, Novartis, and Roche, honoraria from Amgen, BMS, MSD, Novartis, and Roche, travel, accommodation, and other expenses from Amgen, BMS, MSD, Novartis, and Roche, and payment for consultancy or advisory roles from Amgen, BMS, MSD, Novartis, and Roche.

Ethical approval

Ethics approval was obtained at the University of Erlangen-Nürnberg (approval no. 14_13 B dated Feb 19th 2013).

Informed consent

Informed consent was not required as all data were collected as part of routine testing and were fully anonymized prior to analysis.

Supplementary material

428_2018_2480_MOESM1_ESM.docx (38 kb)
ESM 1 (DOCX 37 kb)
428_2018_2480_MOESM2_ESM.docx (35 kb)
ESM 2 (DOCX 35 kb)
428_2018_2480_MOESM3_ESM.docx (30 kb)
ESM 3 (DOCX 30 kb)
428_2018_2480_Fig3_ESM.png (132 kb)

(PNG 132 kb)

428_2018_2480_MOESM4_ESM.eps (2.5 mb)
High Resolution Image (EPS 2517 kb)
428_2018_2480_Fig4_ESM.png (86 kb)

(PNG 86 kb)

428_2018_2480_MOESM5_ESM.eps (2.3 mb)
High Resolution Image (EPS 2369 kb)


  1. 1.
    Garnett MJ, Marais R (2004) Guilty as charged: B-RAF is a human oncogene. Cancer Cell 6:313–319. CrossRefPubMedGoogle Scholar
  2. 2.
    Davies H, Bignell GR, Cox C, Stephens P, Edkins S, Clegg S, Teague J, Woffendin H, Garnett MJ, Bottomley W (2002) Mutations of the BRAF gene in human cancer. Nature 417:949–954. CrossRefPubMedGoogle Scholar
  3. 3.
    Lee JH, Choi JW, Kim YS (2011) Frequencies of BRAF and NRAS mutations are different in histological types and sites of origin of cutaneous melanoma: a meta-analysis. Br J Dermatol 164:776–784. CrossRefPubMedGoogle Scholar
  4. 4.
    Hodis E, Watson IR, Kryukov GV, Arold ST, Imielinski M, Theurillat J, Nickerson E, Auclair D, Li L, Place C (2012) A landscape of driver mutations in melanoma. Cell 150:251–263. CrossRefPubMedGoogle Scholar
  5. 5.
    Jakob JA, Bassett RL Jr, Ng CS, Curry JL, Joseph RW, Alvarado GC, Rohlfs ML, Richard J, Gershenwald JE, Kim KB (2012) NRAS mutation status is an independent prognostic factor in metastatic melanoma. Cancer 118:4014–4023. CrossRefPubMedGoogle Scholar
  6. 6.
    Ehsani L, Cohen C, Fisher KE, Siddiqui MT (2014) BRAF mutations in metastatic malignant melanoma: comparison of molecular analysis and immunohistochemical expression. Appl Immunohistochem Mol Morphol 22:648–651. CrossRefPubMedGoogle Scholar
  7. 7.
    Dummer R, Hauschild A, Lindenblatt N, Pentheroudakis G, Keilholz U, ESMO Guidelines Committee (2015) Cutaneous melanoma: ESMO clinical practice guidelines for diagnosis, treatment and follow-up. Ann Oncol 26(Suppl. 5):v126–v132. CrossRefPubMedGoogle Scholar
  8. 8.
    Chapman PB, Hauschild A, Robert C, Haanen JB, Ascierto P, Larkin J, Dummer R, Garbe C, Testori A, Maio M (2011) Improved survival with vemurafenib in melanoma with BRAF V600E mutation. N Engl J Med 364:2507–2516. CrossRefPubMedGoogle Scholar
  9. 9.
    Hauschild A, Grob JJ, Demidov LV, Jouary T, Gutzmer R, Millward M, Rutkowski P, Blank CU, Miller WH, Kaempgen E (2012) Dabrafenib in BRAF-mutated metastatic melanoma: a multicentre, open-label, phase 3 randomised controlled trial. Lancet 380:358–365. CrossRefPubMedGoogle Scholar
  10. 10.
    Zelboraf® (vemurafenib). Summary of product characteristics. In:. Accessed Feb 2018
  11. 11.
    Tafinlar® (dabrafenib). Summary of product characteristics. In:. Accessed Feb 2018
  12. 12.
    Amanuel B, Grieu F, Kular J, Millward M, Iacopetta B (2012) Incidence of BRAF p.Val600Glu and p.Val600Lys mutations in a consecutive series of 183 metastatic melanoma patients from a high incidence region. Pathology 44:357–359. CrossRefPubMedGoogle Scholar
  13. 13.
    Harlé A, Salleron J, Franczak C, Dubois C, Filhine-Tressarieu P, Leroux A, Merlin JL (2016) Detection of BRAF mutations using a fully automated platform and comparison with high resolution melting, real-time allele specific amplification, immunohistochemistry and next generation sequencing assays, for patients with metastatic melanoma. PLoS One 11:e0153576. CrossRefPubMedGoogle Scholar
  14. 14.
    Long GV, Menzies AM, Nagrial AM, Haydu LE, Hamilton AL, Mann GJ, Hughes TM, Thompson JF, Scolyer RA, Kefford RF (2011) Prognostic and clinicopathologic associations of oncogenic BRAF in metastatic melanoma. J Clin Oncol 29:1239–1246. CrossRefPubMedGoogle Scholar
  15. 15.
    Menzies AM, Haydu LE, Visintin L, Carlino MS, Howle JR, Thompson JF, Kefford RF, Scolyer RA, Long GV (2012) Distinguishing clinicopathologic features of patients with V600E and V600K BRAF-mutant metastatic melanoma. Clin Cancer Res 18:3242–3249. CrossRefPubMedGoogle Scholar
  16. 16.
    Qu K, Pan Q, Zhang X, Rodriguez L, Zhang K, Li H, Ho A, Sanders H, Sferruzza A, Cheng SM, Nguyen D, Jones D, Waldman F (2013) Detection of BRAF V600 mutations in metastatic melanoma: comparison of the Cobas 4800 and Sanger sequencing assays. J Mol Diagn 15:790–795. CrossRefPubMedGoogle Scholar
  17. 17.
    Huang WK, Kuo TT, Wu CE, Cheng HY, Hsieh CH, Hsieh JJ, Shen YC, Hou MM, Hsu T, Chang JW (2016) A comparison of immunohistochemical and molecular methods used for analyzing the BRAF V600E gene mutation in malignant melanoma in Taiwan. Asia Pac J Clin Oncol 12:403–408. CrossRefPubMedGoogle Scholar
  18. 18.
    Colombino M, Capone M, Lissia A, Cossu A, Rubino C, De Giorgi V, Massi D, Fonsatti E, Staibano S, Nappi O, Pagani E, Casula M, Manca A, Sini M, Franco R, Botti G, Caracò C, Mozzillo N, Ascierto PA, Palmieri G (2012) BRAF/NRAS mutation frequencies among primary tumors and metastases in patients with melanoma. J Clin Oncol 30:2522–2529. CrossRefPubMedGoogle Scholar
  19. 19.
    Edmunds SC, Cree IA, Di Nicolantonio F, Hungerford JL, Hurren JS, Kelsell DP (2003) Absence of BRAF gene mutations in uveal melanomas in contrast to cutaneous melanomas. Br J Cancer 88:1403–1405. CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Arndt Hartmann
    • 1
  • Peter Schirmacher
    • 2
  • William Sterlacci
    • 3
  • Winfried Koch
    • 4
  • David B. Liesenfeld
    • 5
  • Birgit Schif
    • 5
  • Claus Garbe
    • 6
  1. 1.Institute of PathologyUniversity of Erlangen-NürembergErlangenGermany
  2. 2.Institute of PathologyUniversity Medicine HeidelbergHeidelbergGermany
  3. 3.Institute of PathologyKlinikum BayreuthBayreuthGermany
  4. 4.BDS KochSchwetzingenGermany
  5. 5.Roche Pharma AGGrenzach-WyhlenGermany
  6. 6.Division of Dermato-OncologyUniversity-Department of DermatologyTübingenGermany

Personalised recommendations