title: Platelet recovery and survival measured in patients by quantitative polymerase chain reaction of mitochondrial DNA.
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.
  
  
  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 × 10(3) 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: r(2)  = 0.957, slope = 0.87).
  
  
  CONCLUSIONS
  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.
authors: DRK-Blutspendedienst NSTOB, Institut Bremen-Oldenburg, Oldenburg, Germany.
authors: Andrea
authors: A
authors: Doescher
authors: Eduard K
authors: EK
authors: Petershofen
authors: Bernd
authors: B
authors: Hertenstein
authors: Doris
authors: D
authors: Kraemer
authors: Jochen
authors: J
authors: Casper
authors: Jörg-Peter
authors: JP
authors: Schmidt
authors: Thomas H
authors: TH
authors: Müller
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000483:Alleles; D001792:Blood Platelets / Q000737:chemistry*; D002470:Cell Survival; D004272:DNA, Mitochondrial / Q000097:blood* / Q000235:genetics; D005260:Female; D005787:Gene Frequency; D005819:Genetic Markers; D006801:Humans; D008297:Male; D008875:Middle Aged; D010865:Pilot Projects; D017713:Platelet Transfusion; D020641:Polymorphism, Single Nucleotide; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D044465:White People / Q000235:genetics
chemical_list: D004272:DNA, Mitochondrial; D005819:Genetic Markers