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Mitochondrial DNA multiplex real-time polymerase chain reaction inhibition assay for quality control of pathogen inactivation by ultraviolet C light in platelet concentrates

Authors
 Sinyoung Kim  ;  Wiebke Handke  ;  Ute Gravemann  ;  Andrea Doscher  ;  Veronika Brixner  ;  Thomas H Muller  ;  Axel Seltsam 
Citation
 Transfusion, Vol.58(3) : 758-765, 2018 
Journal Title
 Transfusion 
ISSN
 0041-1132 
Issue Date
2018
Abstract
BACKGROUND: Several ultraviolet (UV) light-based pathogen inactivation (PI) technologies for platelet (PLT) products have been developed or are under development. Upon implementation of PI technologies, quality control measures are required to ensure consistent efficiency of the treatment process. Previous reports showed that amotosalen/UVA and riboflavin/UV-based PI technologies induce modifications of the PLT-derived mitochondrial DNA (mtDNA) that can be detected by polymerase chain reaction (PCR) inhibition assays. In this study, we sought to establish a PCR inhibition assay to document the impact of ultraviolet C (UVC) treatment with the THERAFLEX UV-Platelets system on the mitochondrial genome in PLT concentrates (PCs). STUDY DESIGN AND METHODS: A multiplex real-time PCR inhibition assay with simultaneous short-amplicon (143 bp) and long-amplicon (794 bp) amplification was developed to detect mtDNA modifications in PLTs after UVC treatment. Assay performance was tested in UVC-treated and untreated, plasma-reduced pooled PCs, and apheresis PCs and challenged using PCs manufactured for a clinical trial under routine-like conditions. RESULTS: UVC illumination of PLTs resulted in dose-dependent inhibition of mtDNA amplification for the larger amplicon. Amplification of the shorter amplicon was not affected by UVC treatment. Evaluation of 283 blinded apheresis and pooled PLT samples from routine-like PC production resulted in prediction of UVC treatment status with 100% accuracy. CONCLUSION: The proposed dual-amplicon size real-time mtDNA PCR assay effectively detects nucleic acid damage induced by UVC illumination of PLTs and could be useful as an informative indicator of PI quality of the THERAFLEX UV-Platelets system.
Full Text
https://onlinelibrary.wiley.com/doi/abs/10.1111/trf.14464
DOI
10.1111/trf.14464
Appears in Collections:
1. Journal Papers (연구논문) > 1. College of Medicine (의과대학) > Dept. of Laboratory Medicine (진단검사의학교실)
Yonsei Authors
김신영(Kim, Sin Young) ORCID logo https://orcid.org/0000-0002-2609-8945
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URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/162149
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