微孔板木瓜酶法筛查不规则抗体技术关键参数的确定  

Determination of key parameter of screening irregular antibody by Papain methods in microplate

作　　者：董珀 许飞[1] 曹锁春[1] 杨爱龙[1] DONG Po;XU Fei;CAO Suochun;YANG Ailong(Changzhou Central Blood Station Testing Center,Changzhou,213000,China)

机构地区：[1]常州市中心血站检测中心,江苏常州213000

出　　处：《临床血液学杂志》2021年第12期874-877,共4页Journal of Clinical Hematology

基　　金：2020年度常州市卫生健康委科技项目(No:WZ202017)。

摘　　要：目的:研究影响微孔板酶法试验的关键参数,确定适合常州市中心血站微孔板酶法检测不规则抗体的最优条件。方法:混合6名献血者的O细胞,鉴定其抗原的多样性,判定其能否作为抗体筛选细胞。以试管木瓜酶法为基础,正交试验为原理,研究不同血浆和酶的量、不同酶的浓度对反应结果的影响,以此确立试验反应体系;并通过比对确定试验关键步骤的震荡参数。结果:6人份混合O细胞符合抗体筛选细胞的要求,选用筛选细胞与血浆、酶液的容量比例为1︰4,标化的酶溶液的浓度为1.25 g/100 mL PBS的反应体系,影响试验关键步骤的震荡最适频率为750 r/min、震荡最适时间为60 s;Lisswell软件读取中,凝集反应可靠扫描点范围是55%。结论:微孔板木瓜酶法筛查不规则抗体技术关键参数可以确定,为大规模筛查不规则抗体奠定了良好的基础。Objective: To research the key parameter that influence the experiment of microplate Papain, and determine the optimal conditions that be suitable for screening irregular antibody of microplate Papain in our blood station. Methods: The antigendiversityof mixedred cells from 6 blood donors were identified as O type, and determined whether they could be used as antibody screening cells. Based on traditional Papain tube method, the influence that different volumes of plasma and enzymes and different concentrations of enzymes on experimental resultswere researched by the orthogonal principle, and the parameters of shocking in key steps were determined through the comparisons. Results: 6-person mixed O cells met the requirements of antibody screening cells. The volume ratio of screening cells to plasma and enzyme was 1︰4 and the concentrations of standardized enzymes were 1.25 g/100 mL PBS in the optimum reaction system. The optimum shocking frequency was 750 r/min and the optimum shocking time was 60 s which influenced the key points of the experiment. The result could beread by Lissweill software, the range of reliable scaning points reflected by agglutination was 55%. Conclusion: The key parameter of screening irregular antibody by Papain methods in microplate can be determined, which may be good base for large-scale routine screening of irregular antibodies.

关 键 词：微孔板 木瓜酶法 不规则抗体筛查 

分 类 号：R457.1[医药卫生—治疗学]