Sharma Praveen, Sachdeva Man Updesh Singh, Kumar Narender, Bose Sunil, Bose Parveen, Uppal Varun, Malhotra Pankaj, Bansal Deepak, Varma Neelam, Ahluwalia Jasmina
Department of Hematology, Advanced Pediatric Centre, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
Clinical Hematology Division, Department of Internal Medicine, Advanced Pediatric Centre, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
Blood Res. 2021 Jun 30;56(2):109-118. doi: 10.5045/br.2021.2020232.
Platelet aggregation studies using conventional light transmission aggregometry (LTA) have several disadvantages and require strict pre-analytical measures for reliable results. We aimed to examine the utility of flow cytometric platelet aggregation (FCA) assay in detecting platelet function defects (PFDs) in patients with a history of bleeding symptoms.
Sixty-four participants (24 patients and 40 healthy controls) were included in this study. LTA and FCA assay were performed simultaneously in patients and healthy controls. In the FCA assay, two portions of platelets from the same individual were labeled separately with CD31-FITC and CD31-PE. After mixing and stimulation with agonists, the double- colored platelet aggregates were visualized using a flow cytometer. The results generated using the two techniques were compared and correlated.
The patients' median age was 17 years (range, 3‒72 yr) with a male-to-female ratio of 1:1.7. There was substantial agreement between LTA and FCA assay in detecting a PFD (=0.792). Four patients showing a Glanzmann thrombasthenia-like pattern on LTA exhibited an abnormal FCA. A functional defect in collagen binding was detected on the FCA assay conducted in two immune thrombocytopenic patients with severe bleeding.
FCA assay can be used to identify functional defects in platelets, with potential applications in thrombocytopenic individuals. It also facilitates the diagnosis of inherited bleeding disorders with platelet defects.
使用传统光透射聚集法(LTA)进行血小板聚集研究存在若干缺点,并且需要严格的分析前措施以获得可靠结果。我们旨在研究流式细胞术血小板聚集(FCA)检测在有出血症状病史患者中检测血小板功能缺陷(PFD)的效用。
本研究纳入64名参与者(24例患者和40名健康对照)。在患者和健康对照中同时进行LTA和FCA检测。在FCA检测中,来自同一个体的两份血小板分别用CD31 - FITC和CD31 - PE标记。混合并用激动剂刺激后,使用流式细胞仪观察双色血小板聚集体。比较并关联两种技术产生的结果。
患者的中位年龄为17岁(范围3 - 72岁),男女比例为1:1.7。在检测PFD方面,LTA和FCA检测之间存在高度一致性(κ = 0.792)。4例在LTA上显示出类似Glanzmann血小板无力症模式的患者FCA异常。在两名有严重出血的免疫性血小板减少症患者进行的FCA检测中检测到胶原结合功能缺陷。
FCA检测可用于识别血小板功能缺陷,在血小板减少个体中有潜在应用。它也有助于诊断伴有血小板缺陷的遗传性出血性疾病。