Chimerism-Recipient Engraftment (Post)
Method Description Describes how the test is performed and provides a method-specific reference
Genomic DNA is extracted from blood or bone marrow aspirate samples using an automated extraction machine (MagNa Pure Compact, Roche Diagnostics) and used in a commercial kit AmpFLSTR Profiler Plus PCR Amplification Kit (Applied Biosystems, Foster City, CA) following the manufacturer's instructions. Briefly, 10 different short tandem repeat (STR) marker regions are amplified in single multiplex PCR using primers labeled with fluorescent tags. The products are analyzed for size and amount using capillary electrophoresis on an ABI3100 genetic analyzer. For the initial sample on any patient, the test is performed on 3 separate DNA samples: donor germline DNA, recipient germline DNA, and recipient posttransplant sample for chimerism determination. The STR profile of the germline samples is used to identify markers that can distinguish between the donor and recipient. Based on these profiles, the percentage of donor and recipient DNA is then determined in the posttransplant sample using the assumptions and calculations outlined in Thiede, et al. Subsequent samples for chimerism evaluation do not need to be accompanied by samples for donor and recipient germline evaluation, as the profiles from the initial testing are kept on file for comparison.(MagNA Pure Compact System. User Manual 2004. Roche Diagnostics, Basel, Switzerland. www.roche-applied-science.com; AmpFLSTR Profiler Plus PCR Amplification Kit. User Bulletin 2004. Applied Biosystems, Foster City, CA. www.appliedbiosystems.com; Thiede C, Florek M, Bornhauser M, et al: Rapid quantification of mixed chimerism using multiplex amplification of short tandem repeat markers and fluorescence detection. Bone Marrow Transplant 1999;23:1055-1060)
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; 3 p.m.