Cytochrome P450 2D6 (CYP2D6) Comprehensive Cascade, Blood
Method Description Describes how the test is performed and provides a method-specific reference
Genomic DNA is extracted from whole blood.
Luminex Assay (tier 1): The xTAG assay for CYP2D6 incorporates multiplex PCR and multiplex allele-specific primer extension (ASPE) with Luminex Molecular Diagnostics' proprietary universal tag sorting system on the Luminex 100 xMAP platform.(Package insert: xTAG Kit for CYP2D6, Luminex Molecular Diagnostics Inc, Austin, TX)
Copy Number Assay (tier 2): This assay utilizes a duplex real-time PCR, which includes 1 copy number probe (FAM labeled) and a reference assay (VIC labeled) per reaction. Each copy number probe detects the genomic sequence of interest and the reference assay detects a sequence that is known to be present in 2 copies in a diploid genome. Relative quantitation is used to determine the relative copy number of the target of interest in a genomic DNA (gDNA) sample normalized to 10 ng/mcL for each probe. Each probe is normalized to the known copy number of the reference sequence, and compared to a calibrator sample with known copies of the target sequence included with each run.(Package insert: Taqman Copy Number Assays Revision D, Applied Biosystems, Carlsbad, CA)
Sequencing component assays (tier 3): The CYP2D6 gene of interest is amplified by PCR. The PCR product is then purified and sequenced in both directions using fluorescent dye-terminator chemistry. Sequencing products are separated on an automated sequencer and trace files analyzed for variations in the exons and intron/exon boundaries of all 9 exons using mutation detection software and visual inspection.(Unpublished Mayo method)
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.
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
Whole Blood: 2 weeks Extracted DNA: 2 months
Performing Laboratory Location The location of the laboratory that performs the test