BRAF Mutation (T1799A) Analysis by PCR and Sequencing, Thyroid
Method Description Describes how the test is performed and provides a method-specific reference
Tissue from either cytology slides or paraffin-embedded tissue is lysed and digested. Genomic DNA is extracted from thyroid specimens using either a phenol-chloroform method or the QIAamp DNA FFPE Tissue kit (Qiagen). The DNA is amplified via PCR. Primers specific for the BRAF exon 15 gene are used. Controls are run with each specimen to assess possible contamination issues and overall test performance. The patient and wild-type samples are sent for direct DNA sequencing. The sequencing chromatograms are analyzed by manual and software methods and the presence or absence of the BRAF mutation (T1799A) is determined. The results are interpreted and reported by a working group pathologist.(Jin L, Sebo TJ, Nakamura N, et al: BRAF mutation analysis in fine needle aspiration (FNA) cytology of the thyroid. Diagn Mol Pathol 2006;15:136-143; Nakamura N, Carney JA, Jin L, et al: RASSF1A and NORE1A methylation and BRAFV600E mutations in thyroid tumors. Lab Invest 2005;85:1065-1075)
Day(s) and Time(s) Test Performed Outlines the days and times the test is performed. This field reflects the day and time the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time required before the test is performed. Some tests are listed as continuously performed, which means assays are performed several times during the day.
Monday through Friday; 8 a.m.–5 p.m.
Analytic Time Defines the amount of time it takes the laboratory to setup and perform the test. This is defined in number of days. The shortest interval of time expressed is "same day/1 day," which means the results may be available the same day that the sample is received in the testing laboratory. One day means results are available 1 day after the sample is received in the laboratory.
Maximum Laboratory Time Defines the maximum time from specimen receipt at Mayo Medical Laboratories until the release of the test result
Specimen Retention Time Outlines the length of time after testing that a specimen is kept in the laboratory before it is discarded
1 week/7 days
Performing Laboratory Location The location of the laboratory that performs the test