BCR/ABL, p210, mRNA Detection, Reverse Transcription-PCR (RT-PCR), Quantitative, Monitoring Chronic Myelogenous Leukemia (CML)
Method Description Describes how the test is performed and provides a method-specific reference
The assay is performed using an automated platform, GeneXpert (Cepheid). A buffy coat is made from whole blood and 200 microliters, or a maximum of 10 million cells, are added to a single sample cartridge and loaded onto the GeneXpert machine. All subsequent reactions are performed within the cartridge and the results are calculated by the instrument. Within the cartridge, RNA is extracted and converted to complementary DNA (cDNA). Quantitative, reverse transcription PCR is performed with a nested PCR reaction containing primers designed to amplify cDNA from the e13/a2 and e14/a2 bcr/abl fusion products. A fragment of abl cDNA is also amplified as a control for RNA degradation. The ratio of bcr/abl(p210) to abl is calculated from the difference in the crossing thresholds of bcr/abl(p210) and abl products. Lot-to-lot variation in the cartridges is corrected using a calibrator evaluated at the manufacturer.(Unpublished Mayo method)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