用于诊断 Glanzmann 血小板无力症和杂合子携带者的新型 ELISA 方法。

A novel ELISA for diagnosis of Glanzmann's thrombasthenia and the heterozygote carriers.

机构信息

National Institute of Immunohematology, KEM. Hospital Campus, Parel, Mumbai, India.

出版信息

Ann Hematol. 2012 Jun;91(6):917-21. doi: 10.1007/s00277-011-1390-1. Epub 2011 Dec 21.

Abstract

A sensitive and specific sandwich ELISA was developed for the diagnosis of Glanzmann's thrombasthenia (GT) and the heterozygote carriers of the disease using whole blood platelets. The assay used anti-CD36 antibody to capture platelets from platelet-rich plasma which was subsequently treated with a bioengineered disintegrin/alkaline phosphatase hybrid protein specific for GP IIb/IIIa. The test allows large number of samples to be typed and can also be used on stored samples. The assay correctly diagnosed 40 normal healthy individuals, 10 GT cases, 10 heterozygotes, 3 Bernard-Soulier syndrome cases and 2 type 3 GT cases. ELISA plates were stable at room temperature up to 3 weeks without any loss of activity. This novel and simple test can be widely used for heterozygote detection besides diagnosing GT cases without using a sophisticated flow cytometer or a platelet aggregometer and has wide applicability in countries like India where many of these cases remain undiagnosed due to the lack of diagnostic facilities.

摘要

一种敏感且特异的夹心 ELISA 法被开发出来，用于诊断 Glanzmann 血小板无力症（GT）和疾病的杂合子携带者，使用的是全血血小板。该检测方法使用抗 CD36 抗体从富含血小板的血浆中捕获血小板，随后用针对 GP IIb/IIIa 的生物工程整联蛋白/碱性磷酸酶杂合蛋白处理。该检测方法可以对大量样本进行分型，也可以用于储存样本。该检测方法正确诊断了 40 名正常健康个体、10 例 GT 病例、10 例杂合子、3 例 Bernard-Soulier 综合征病例和 2 例 3 型 GT 病例。ELISA 板在室温下稳定 3 周，没有任何活性损失。这种新颖且简单的检测方法除了诊断 GT 病例外，还可以广泛用于杂合子检测，而无需使用复杂的流式细胞仪或血小板聚集仪，在印度等许多这些病例由于缺乏诊断设施而未被诊断的国家具有广泛的适用性。