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数字聚合酶链反应监测心脏手术患者的血小板输注情况。

Digital polymerase chain reaction to monitor platelet transfusions in cardiac surgery patients.

作者信息

Ali Khaldoun, Müller Thomas H, Garritsen Henk S P, Harringer Wolfgang, Doescher Andrea

机构信息

Clinic for Cardiac, Thoracic and Vascular Surgery, Klinikum Braunschweig GmbH, Braunschweig, Germany.

Institute for Clinical Transfusion Medicine, Klinikum Braunschweig GmbH, Braunschweig, Germany.

出版信息

Vox Sang. 2023 May;118(5):384-391. doi: 10.1111/vox.13422. Epub 2023 Mar 13.

Abstract

BACKGROUND AND OBJECTIVES

Corrected count increment (CCI) measurements monitor the effectiveness of platelet transfusions in haemato-oncology, but they usually fail in patients undergoing cardiac surgery. We investigated whether polymerase chain reaction (PCR) of mitochondrial single-nucleotide polymorphisms (SNPs) is able to monitor the survival of transfused platelets in these patients.

MATERIALS AND METHODS

Leukocyte-free, platelet-rich plasma was prepared from patients' blood to measure platelet counts based on patient-/donor-specific SNPs by digital PCR after DNA extraction. Platelet counts in samples from patients with severe thrombocytopenia were analysed by both PCR and flow cytometry. Ten patients undergoing cardiac surgery with the use of heart lung machine and without overt bleeding received a single apheresis platelet concentrate because of either dual platelet inhibition during a non-elective intervention or a complex procedure. Blood samples were collected at nine defined intervals (0-120 h) post transfusion.

RESULTS

The digital PCR of the seven SNPs reliably quantified levels ≥0.6 G/L platelets, in good agreement with flow cytometry and without interference by other SNPs or by platelet activation. A mean 24-h CCI of 11.8 (range: 5.6-19.8) and a mean 120-h area under the curve (AUC) of 1386 (915-1821) hxG/L were observed for the transfused platelets. The mean AUC of 14,103 (3415-27,305) hxG/L for the patients' endogenous platelets indicates that transfused platelets represented only 11% (5-25) of the total platelet counts during 120 h post transfusion.

CONCLUSION

PCR of mitochondrial SNPs offers a tool to assess the survival of platelets from apheresis concentrates in cardiac surgery patients to facilitate the implementation of improved transfusion strategies.

摘要

背景与目的

校正计数增加值(CCI)测量可监测血液肿瘤学中血小板输注的有效性,但在心脏手术患者中通常效果不佳。我们研究了线粒体单核苷酸多态性(SNP)的聚合酶链反应(PCR)是否能够监测这些患者中输注血小板的存活情况。

材料与方法

从患者血液中制备无白细胞的富血小板血浆,DNA提取后通过数字PCR基于患者/供体特异性SNP测量血小板计数。对严重血小板减少症患者样本中的血小板计数进行PCR和流式细胞术分析。10例使用心肺机且无明显出血的心脏手术患者,因非选择性干预期间的双重血小板抑制或复杂手术,接受了单次单采血小板浓缩物输注。在输血后9个规定时间间隔(0 - 120小时)采集血样。

结果

7个SNP的数字PCR可靠地定量了≥0.6 G/L血小板水平,与流式细胞术结果高度一致,且不受其他SNP或血小板激活的干扰。输注血小板的平均24小时CCI为11.8(范围:5.6 - 19.8),平均120小时曲线下面积(AUC)为1386(915 - 1821)hxG/L。患者内源性血小板的平均AUC为14,103(3415 - 27,305)hxG/L,表明输注的血小板在输血后120小时内仅占总血小板计数的11%(5 - 25)。

结论

线粒体SNP的PCR为评估心脏手术患者单采浓缩物中血小板的存活情况提供了一种工具,有助于实施改进的输血策略。

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