Comparison of automated short tandem repeat and manual variable number of tandem repeat analysis of chimerism in bone marrow transplant patients.
Hematopoietic chimerism can be monitored in bone marrow transplant patients at DNA polymorphic sites. In this study, allele detection and quantification by ethidium bromide-stained agarose gels were compared with automated fluorescent sizing on an artificially mixed system and on chimeric post-transplant whole blood and sorted cell populations. A panel of five variable number of tandem repeats (VNTRs) were amplified and quantified visually on an ethidium bromide-stained gel. The ten short tandem repeats (STRs) were amplified as a multiplex polymerase chain reaction (PCR) and fluorescently detected on a DNA sequencer. Fluorescent band intensities were converted to fluorescent peak areas for allele quantification. Using mixed DNA of different proportions, both STRs and VNTRs showed linearity and appeared equally sensitive. However, case studies showed STRs to be more sensitive (<5%) than VNTRs (<10%). The STRs more accurately quantified the minor DNA component at low concentrations.
Millson
A S
AS
Associated Regional and University Pathologists, Inc. (ARUP Laboratories), Salt Lake City, Utah 84108, USA.
Spangler
F L
FL
Wittwer
C T
CT
Lyon
E
E
D000483:Alleles; D001322:Autoanalysis / Q000191:economics / Q000379:methods; D001854:Bone Marrow Cells; D016026:Bone Marrow Transplantation; D004247:DNA / Q000032:analysis; D004587:Electrophoresis, Agar Gel / Q000191:economics / Q000379:methods; D006801:Humans; D018598:Minisatellite Repeats / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D018183:Transplantation Chimera / Q000235:genetics*
D004247:DNA