BCR/ABL, p190, mRNA Detection, Reverse Transcription-PCR (RT-PCR), Quantitative, Monitoring Assay
Method Description Describes how the test is performed and provides a method-specific reference
Total RNA is extracted from the sample using the QIAmp kit (Qiagen). cDNA is made using SuperScript III Kit (Invitrogen). A quantitative, real-time-PCR reaction is performed using the LightCycler 2.0 instrument (Roche) with one set of primers designed to detect the bcr/abl e1/e2 fusion and a second set of primers designed to detect a fragment of abl. Taqman-type probe technology is used in both reactions. The data is analyzed using the supplied software for relative quantification with calibrator normalization and efficiency correction. (Roche Applied Science LightCycler 2.0 Instrument Operation Manual, Version 4/1.2. 2003). The abl is amplified to control for RNA degradation in the sample and the calibrator is used to control for inter-run variations. A normalized ratio of bcr/abl(p190) mRNA:abl mRNA is obtained and converted to the reported value of percent bcr/abl(p190):abl.(Unpublished Mayo method)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.
Monday through Friday a.m.