Platelet recovery and survival measured in patients by quantitative polymerase chain reaction of mitochondrial DNA
Version of Record online: 23 JUL 2014
© 2014 AABB
Volume 55, Issue 1, pages 55–63, January 2015
How to Cite
Doescher, A., Petershofen, E. K., Hertenstein, B., Kraemer, D., Casper, J., Schmidt, J.-P. and Müller, T. H. (2015), Platelet recovery and survival measured in patients by quantitative polymerase chain reaction of mitochondrial DNA. Transfusion, 55: 55–63. doi: 10.1111/trf.12778
- Issue online: 13 JAN 2015
- Version of Record online: 23 JUL 2014
- Manuscript Accepted: 31 MAY 2014
- Manuscript Revised: 27 MAY 2014
- Manuscript Received: 3 MAR 2014
Mitochondrial (mt) DNA markers have been identified as potential targets for the quantification of endogenous and allogeneic platelets (PLTs) in the blood of individuals who received transfusions. Our goal was to develop a routine polymerase chain reaction (PCR) assay for ex vivo monitoring of PLT survival in patients after transfusion.
Study Design and Methods
Targets were selected for real-time (RT)-PCR of mt DNA based on the frequency distribution of nucleotide polymorphisms and assay sensitivity in vitro. The assays were then evaluated with ex vivo samples to measure PLT survival and recovery of therapeutic doses of apheresis PLTs in hematooncologic patients with thrombocytopenia.
Nucleotides in two positions (73/310 hypervariable region [HVR] 2) and three positions (295 HVR 2, 16069/16311 HVR 1) had allele frequencies of approximately 0.5 and 0.85, respectively, in a population of 960 Caucasian PLT donors. They provided targets for sensitive assays detecting at least 1 × 103 PLTs per whole blood sample with adequate reproducibility (interassay coefficient of variation <4.0%). Transfusions of single-donor PLT concentrates in patients with thrombocytopenia (n = 30) were monitored with these markers. The mean 24-hour corrected count increment was 8.3 and the mean calculated survival time was 3.3 days. Results for a second marker were available for 13 transfusions. The survival time values derived from both markers for the same transfusion were almost identical (linear regression: r2 = 0.957, slope = 0.87).
This RT-PCR method detects mt DNA polymorphisms in Caucasians for a highly sensitive and reproducible quantification of endogenous and allogeneic PLT numbers in blood samples from transfused patients with thrombocytopenia.