Comparison of In-House Methods and Cobas BRAF V600 Mutation Assay in Melanoma Tumor Samples

This study has been completed.
Sponsor:
Information provided by (Responsible Party):
Hoffmann-La Roche
ClinicalTrials.gov Identifier:
NCT01744860
First received: December 6, 2012
Last updated: February 25, 2016
Last verified: November 2015
  Purpose
This non-interventional study will compare the Cobas BRAF V600 mutation assay with in-house methods used in molecular laboratories for the assessment of the BRAF mutation status in melanoma tumor samples. No patients will be enrolled in this study. Data will be collected for approximately 6 months.

Condition
Malignant Melanoma

Study Type: Observational
Study Design: Observational Model: Cohort
Time Perspective: Prospective
Official Title: Evaluation of Concordance Between the Methods Used in INCa Platforms and the Cobas® 4800 BRAF V600 Mutation Test for Detection of BRAF V600 Mutations in Melanoma in Real Life Setting

Resource links provided by NLM:


Further study details as provided by Hoffmann-La Roche:

Primary Outcome Measures:
  • BRAF Mutation Status According to Cobas 4800 BRAF V600 Mutation Test vs. INCa Laboratories Molecular Genetics Laboratories [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    BRAF V600 mutation status was determined by INCa molecular laboratories "in-house" methods and Cobas 4800 BRAF V600 mutation test. Samples were analysed as V600 mutation, No V600 mutation and Non evaluable. Additionally, the type of V600 mutation (E, K, R, D, E2, other V600 mutation, not specified) was also evaluated only by INCa molecular laboratory "in-house" method.


Secondary Outcome Measures:
  • Tumor Sample Characteristics-Type of Tumor Sample [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    The type of tumor sample used for evaluation of BRAF V600 mutation whether it was a biopsy or surgical specimen were reported

  • Tumor Sample Characteristics - Source of Tumor Sample [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    The source of tumor sample for BRAF V600 mutation detection whether taken from internal or external pathology laboratory were reported

  • Type of Pathology Laboratory Performing the Fixation or Embedding-Pre-analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    The external or internal pathology laboratories involved in the process of fixation or embedding of the tumor sample was evaluated.

  • Time From Sampling to Fixation- Pre-analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    Time taken from the sampling to the fixation of the tumor sample was reported in range of 0-2 hours, 2-6 hours, >6 hours and unknown. Number of samples falling in each of the class were reported.

  • Type of Fixative Used- Pre-analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    The different types of fixative Excell, formol, alcohol formol acetic acid and other, used to fix the tumor samples were reported.

  • Fixation Duration by Pre-analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    Fixation duration is defined as the amount of time required in hours for the fixation of a samples. The fixation duration was categorized as <6 hours, 6-24 hours and >24 hours and unknown. Number of samples falling in each category were reported

  • Slice Thickness by Pre-analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    Slice thickness of all the tumour samples was measured. The slice thickness was measured in micrometer.

  • Dewaxing by Pre-analytical Method [ Time Frame: Up to 6 Months ] [ Designated as safety issue: No ]
    Dewaxing is a method to recover the DNA from samples. Dewaxing information was collected as "Yes, No or Missing"

  • Necrosis Percentage Determination by Pre-analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    The percentage of necrosis defined as the death of one or more cells in the analysed zone was reported.

  • Percentage of Tumor Cells by Pre-analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    The percentage of tumor cells in the given tumor sample were reported.

  • Tumor Samples With Presence of Melanin by Pre-analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    The tumor samples with presence of melanin were categorized as Important, Few, Medium and Absent.

  • DNA Extraction - Extraction Method by Pre-analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    This method assessed DNA from the tumor samples was extracted by Automated method or Manual method.

  • Median DNA Elution Volume by Pre-analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    Median DNA elution volume microliters [mcl] was reported.

  • Mean DNA Concentration by Pre-analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    The DNA concentration in the tissue elute was measured in nanogram per microliter (ng/mcL).

  • Amount of DNA by Pre-analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    The total DNA concentration extracted from the tissue was measured in nanogram (ng).

  • Size of Amplicons Used by "In-house" Analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    The method described the size of amplicon used. It was measured in base pairs (bp).

  • Method of Mutation Detection by "In-house" Analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    Allele-specific PCR, High Resolution Melting (HRM) + Sanger sequencing, Pyrosequencing, Sanger sequencing, Real time PCR, SNaPshot were used for BRAF V600 mutation detection.

  • Number of Samples Punched in In-house Analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    Total number of samples for whom punch was used in 'in-house analytical' method are reported.

  • Mean Number of Slices Per Sample Used for "In-house"- Analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    The mean of number of slices per sample when no punch was used are reported.

  • Median Time Between Receipt of Samples and Determination of Result by "In-house" Analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    This In-house analytical method measured the time between receipt of samples to the result determination. It measured the time in days.

  • Technician Work Time Between DNA Extraction and Result by "In-house" Analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    The working time required by the technician from the time of DNA extraction to the time to obtain the results was measured in hours.

  • Mean DNA Concentration as Measured by COBAS 4800 BRAF V600 Mutation Test-Analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    The DNA concentration as assessed by COBAS 4800 BRAF V600 Mutation assay was reported. The unit used to measure the DNA concentration was nanogram/microlitre (ng/mcl)

  • Punch Used for Cobas 4800 BRAF V600 Mutation Test- Analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    The punch done during Cobas 4800 BRAF V600 Mutation Test on the sample was described as Yes or No.

  • Number of Slices Used When No Punch Was Used for Cobas 4800 BRAF V600 Mutation Test- Analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    This describes the Cobas 4800 BRAF V600 Mutation Test, for the mean of number of slices when No punch method, was used. Of the 420 samples, punch was Yes, for 45 samples and punch was No, for 375 samples.

  • Median Time Between Receipt of Sample and Determination of Result by Cobas 4800 BRAF V600 Mutation Test -Analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    This analytical method for cobas 4800 BRAF V600 Mutation Test measured the time between receipt of samples to the result determination. It measured the time in days.

  • Technician Work Time Between DNA Extraction and Result by Cobas 4800 BRAF V600 Mutation Test - Analytical Method [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    This cobas 4800 BRAF V600 Mutation Test analytical method measures the working time required by the technician from the time of DNA extraction to the time to obtain the results. The time duration was measured in hours.

  • Management of Discordance- Method Used to Manage Discordance [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    Crossing DNA, DNA from In-House method analysed with cobas, SNaPshot, DNA from cobas analysed with In-House method, external site control test, Sanger sequencing, Kit CE-IVD Therascreen RGQ Qiagen, Kit Therascreen RGQ BRAF + Pyrosequencing by another platform (PF), Pyrosequencing, Mutation detection On Another Block, (primitive tumor [prm. tmr]), Sequencing And Therascreen kit (Qiagen) were used for management of discordance between "in-house" method and Cobas 4800 mutation test.

  • Management of Discordance-Final Result for BRAF V600 Mutation Detection [ Time Frame: Up to 6 months ] [ Designated as safety issue: No ]
    The final results obtained by discordance management of the 28 discordant samples were BRAF V600 mutation, No BRAF V600 mutation and Non-evaluable. These results were further assessed by the Investigator and interpreted as final result.


Enrollment: 420
Study Start Date: December 2012
Study Completion Date: April 2013
Primary Completion Date: April 2013 (Final data collection date for primary outcome measure)
Groups/Cohorts
INCa molecular genetics laboratory "in-house" methods
BRAF V600 mutations were analysed using INCa (Institut National du Cancer [French National Cancer Institute]) molecular genetics laboratories using "in-house" methods
Cobas 4800 Mutation Test
BRAF V600 mutations were analysed using Cobas 4800 mutation test

  Eligibility

Ages Eligible for Study:   Child, Adult, Senior
Genders Eligible for Study:   Both
Accepts Healthy Volunteers:   No
Sampling Method:   Probability Sample
Study Population
No patients are enrolled in this study. Use of melanoma tumor samples.
Criteria

Inclusion Criteria:

No patients are enrolled. Use of tumor samples only.

  • Histologically proven melanoma tumor sample
  • Any type of tumor sample: biopsy or surgical specimen of primary tumor or metastasis
  • Tumor samples must be fixed and paraffin-embedded.

Exclusion Criteria:

No patients are enrolled. Use of tumor samples only.

  • Fixative unknown
  Contacts and Locations
Choosing to participate in a study is an important personal decision. Talk with your doctor and family members or friends about deciding to join a study. To learn more about this study, you or your doctor may contact the study research staff using the Contacts provided below. For general information, see Learn About Clinical Studies.

Please refer to this study by its ClinicalTrials.gov identifier: NCT01744860

Locations
France
Boulogne Billancourt, France, 92104
Colmar, France, 68024
Lille, France, 59037
Lyon, France, 69437
Marseille, France, 13015
Montpellier, France, 34295
Nantes, France, 44093
Paris, France, 75010
Pessac, France, 33604
Rouen, France, 76031
Vandoeuvre Les Nancy, France, 54511
Villejuif, France, 94505
Sponsors and Collaborators
Hoffmann-La Roche
Investigators
Study Director: Clinical Trials Hoffmann-La Roche
  More Information

Responsible Party: Hoffmann-La Roche
ClinicalTrials.gov Identifier: NCT01744860     History of Changes
Other Study ID Numbers: ML28471 
Study First Received: December 6, 2012
Results First Received: January 21, 2016
Last Updated: February 25, 2016
Health Authority: France: Ministry of Health

Additional relevant MeSH terms:
Melanoma
Neuroendocrine Tumors
Neuroectodermal Tumors
Neoplasms, Germ Cell and Embryonal
Neoplasms by Histologic Type
Neoplasms
Neoplasms, Nerve Tissue
Nevi and Melanomas

ClinicalTrials.gov processed this record on July 21, 2016