Platelet recovery and survival measured in patients by quantitative polymerase chain reaction of mitochondrial DNA
- 23 July 2014
- journal article
- clinical trial
- Published by Wiley in Transfusion
- Vol. 55 (1) , 55-63
Abstract
Background: 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.Results: 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 2 = 0.957, slope = 0.87).Conclusion: 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.Keywords
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