In vivo tracking of transfused platelets by next-generation sequencing of mitochondrial DNA

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Authors: Seung Jun Choi ; Soon Sung Kwon ; Hyun Kyung Kim ; Eun Jung Suh ; Haerim Chung ; Hyunsoo Cho ; June-Won Cheong ; Sinyoung Kim

Keywords: Blood Component Removal ; Blood Donors ; Blood Platelets* / cytology ; Blood Platelets* / metabolism ; Cell Separation* / methods ; DNA, Mitochondrial* / analysis ; DNA, Mitochondrial* / genetics ; DNA, Mitochondrial* / metabolism ; Feasibility Studies ; Hematologic Neoplasms / therapy ; High-Throughput Nucleotide Sequencing ; Humans ; Platelet Transfusion* ; Polymorphism, Single Nucleotide ; Treatment Outcome

DOI: Tracking transfused platelets is important to evaluate platelet transfusion efficiency. Traditionally, corrected count increments were used; however, quantitative polymerase chain reaction-based methods have recently been developed. As both these methods have some limitations, we developed a new method based on next-generation sequencing (NGS) of platelet mitochondrial DNA (mtDNA). We identified several single nucleotide variant markers by sequencing the entire mtDNA region of platelets, and used NGS to estimate the proportion of each platelet unit. This method was validated using mixed platelets obtained from different donors at various ratios. We confirmed the applicability of this method in patients who received platelet transfusions using pre- and posttransfusion samples from 12 patients with hematological malignancies. The method showed good linearity (r2 >0.99 in the range of mixing ratios from 1:1 to 1:50) in the platelet-mixing experiment. In addition, platelet tracking in patients who received transfusions was feasible using this method. Furthermore, it was possible to track individual platelets in patients who received a single platelet transfusion and in those who received multiple transfusions, including a patient who received 5 platelet transfusions. Hence, this NGS-based platelet-tracking method can be used for patients with various conditions.

Appears in Collections:: 1. College of Medicine (의과대학) > Dept. of Laboratory Medicine (진단검사의학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Internal Medicine (내과학교실) > 1. Journal Papers

Yonsei Authors: Kwon, Soon Sung(권순성)
Kim, Sin Young(김신영) https://orcid.org/0000-0002-2609-8945
Cheong, June-Won(정준원) https://orcid.org/0000-0002-1744-0921
Chung, Hae Rim(정해림) https://orcid.org/0000-0002-7926-9285
Cho, Hyunsoo(조현수) https://orcid.org/0000-0003-2651-6403
Choi, Seung Jun(최승준) https://orcid.org/0000-0003-0736-1055

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