Hamada Kisei, Koreeda Ako, Ohtsu Yuki, Yonemitsu Kosei, Tsunenari Shigeyuki
Forensic Science Laboratory, Kumamoto Prefectural Police Headquarters, Japan.
Leg Med (Tokyo). 2002 Dec;4(4):217-22. doi: 10.1016/s1344-6223(02)00048-2.
A simple dot enzyme-linked immunosorbent assay (Dot-ELISA) using commercially available monoclonal anti-A and anti-B antibodies and biotinylated anti-H lectin was developed for ABO blood typing of biological fluid and stains. Its application to forensic practice was examined with 117 saliva samples and their stains, and practical case samples of 8 seminal, 6 vaginal and 45 aged salivary stains. In the simple Dot-ELISA, a new step to heat biological samples was introduced in the system in order to block unfavorable non-specific reactions of the samples with secondary enzyme conjugate. The simple Dot-ELISA could determine accurately the ABO blood type of a small amount of secretor's and non-secretor's salivary samples. In practical tests of seminal, vaginal and salivary stains, all results were confirmed to be identical to those determined by the conventional absorption-inhibition test and the absorption-elution test. The simple Dot-ELISA is considered to be accurate, rapid, simple, sensitive and easy to perform in routine forensic practice. It is also a unique and helpful method to determine the ABO blood types of various biological samples.
利用市售抗A和抗B单克隆抗体以及生物素化抗H凝集素开发了一种简单的斑点酶联免疫吸附测定法（Dot-ELISA），用于生物体液和污渍的ABO血型鉴定。用117份唾液样本及其污渍以及8份精液、6份阴道分泌物和45份陈旧唾液污渍的实际案例样本对其在法医实践中的应用进行了检验。在简单的Dot-ELISA中，系统中引入了一个加热生物样本的新步骤，以阻断样本与二级酶结合物的不良非特异性反应。简单的Dot-ELISA能够准确测定少量分泌型和非分泌型唾液样本的ABO血型。在精液、阴道分泌物和唾液污渍的实际测试中，所有结果均被证实与传统吸收抑制试验和吸收洗脱试验所确定的结果一致。简单的Dot-ELISA被认为在常规法医实践中准确、快速、简单、灵敏且易于操作。它也是一种用于确定各种生物样本ABO血型的独特且有用的方法。
