Jung Bo Kyeung, Cho Chi Hyun, Moon Kyung Chul, Sung Hur Dae, Yoon Jeong-Ah, Yoon Soo-Young
1. Department of Laboratory Medicine, College of Medicine, Korea University Guro Hospital, Seoul 152-703, South Korea.
2. Nanoentek Incorp., Seoul, Korea.
Int J Med Sci. 2014 Sep 18;11(12):1228-33. doi: 10.7150/ijms.10008. eCollection 2014.
Inappropriate platelet activation is known to be associated with various thrombotic disorders. Platelet-monocyte aggregates (PMAs), whose formation is mediated by platelet surface P-selectin (CD62P), can be used as a reliable marker to detect platelet activation. Previous studies have generally detected PMAs through flow cytometry-based approaches. Recently, the ADAM(®) image cytometer (Nanoentek Inc., Seoul, Korea) was developed for image-based cellular analysis. In this study, we detected PMAs with the ADAM(®) cytometer, evaluated the reproducibility of the measurements made by the ADAM(®) cytometer, and compared the abilities of the ADAM(®) cytometer and a flow cytometric assay to detect PMAs.
Whole blood samples were collected from patients. Within 5 minutes of collection, anticoagulated whole blood samples were fixed in 10% paraformaldehyde and 5% glyoxal. Nineteen clinical specimens were collected; each was analyzed three times with the ADAM(®) cytometer in order to assess the reproducibility of its measurements. To compare the ability of the ADAM(®) cytometer with that of a flow cytometer to detect PMAs, each cytometer was used for 23 clinical samples and the correlation of the measurements was determined.
The PMA measurements made by the ADAM(®) cytometer showed good reproducibility (CV < 10% for all specimens). Moreover, the PMA measurements made by the ADAM(®) cytometer exhibited a high correlation with those made by a flow cytometric assay (R = 0.944).
The ADAM(®) cytometer is a suitable alternative method to the flow cytometry-based assays. Since the ADAM cytometer does not need specialized instrument knowledge or software proficiency (unlike flow cytometry), the ADAM(®) cytometer can be used as a rapid and reliable POCT device to measure platelet activation in peripheral blood. This, in turn, will provide valuable information regarding patient propensities to thrombotic diseases.
已知不适当的血小板激活与多种血栓性疾病相关。血小板 - 单核细胞聚集体（PMA）的形成由血小板表面P - 选择素（CD62P）介导，可作为检测血小板激活的可靠标志物。以往研究通常通过基于流式细胞术的方法检测PMA。最近，ADAM（®）图像细胞仪（韩国首尔Nanoentek公司）被开发用于基于图像的细胞分析。在本研究中，我们使用ADAM（®）细胞仪检测PMA，评估ADAM（®）细胞仪测量的可重复性，并比较ADAM（®）细胞仪和流式细胞术检测PMA的能力。
从患者采集全血样本。采集后5分钟内，将抗凝全血样本固定于10%多聚甲醛和5%乙二醛中。收集了19份临床标本；每份标本用ADAM（®）细胞仪分析三次，以评估其测量的可重复性。为比较ADAM（®）细胞仪与流式细胞仪检测PMA的能力，每种细胞仪用于检测23份临床样本，并确定测量结果的相关性。
ADAM（®）细胞仪进行的PMA测量显示出良好的可重复性（所有标本的CV < 10%）。此外，ADAM（®）细胞仪进行的PMA测量与流式细胞术测量结果高度相关（R = 0.944）。
ADAM（®）细胞仪是基于流式细胞术检测方法的合适替代方法。由于ADAM细胞仪不需要专门的仪器知识或软件操作能力（与流式细胞术不同），ADAM（®）细胞仪可作为一种快速可靠的即时检验设备，用于测量外周血中的血小板激活情况。这反过来将为患者发生血栓性疾病的倾向提供有价值的信息。
