流式荧光法检测抗MICB抗体的建立及初步应用  

Development and application of the method for detection of anti-MICB antibodies by using Luminex flow cytometry

作　　者：樊一笋[1] 曾万杰[1] 耿春松[1] 胡振华[1] 陈强[1] 丁双双[1] 

机构地区：[1]上海交通大学医学院苏州九龙医院检验科,江苏苏州215021

出　　处：《临床检验杂志》2011年第8期567-569,共3页Chinese Journal of Clinical Laboratory Science

摘　　要：目的建立稳定、快速和特异的抗MHC-I类链相关基因B(MICB)抗体流式荧光检测方法,了解MICB抗体在健康人群中的分布频率。方法克隆、表达和分离纯化MICB融合蛋白,将其包被在Luminex微球表面,建立MICB抗体流式荧光检测方法。评价方法的线性范围、特异性、精密度、准确性,并用此方法检测500例健康人血清MICB抗体。结果 MICB抗体流式荧光检测方法在血清抗体浓度在3.1～100.0μg/mL之间,具有较好的线性关系,相关系数为0.995。回收实验和重复实验结果显示,其回收率为97.4%～99.6%,批内变异系数为3.4%、2.9%,批间变异系数为4.5%、4.9%。该检测方法与MICA抗体无交叉反应。500例健康人血清中发现4例MICB等位基因特异性抗体。结论 MICB抗体流式荧光检测方法的特异性和灵敏度较高,结果稳定可靠。MICB抗体在器官移植中的作用需进一步研究。Objective To develop a multiplex flow method for the detection of antibodies against MICB（major histocompatibility complex class I chain related molecule B）,and analyze the distribution frequency of MICB antibodies in healthy adults.Methods Recombinant proteins of MICB were produced by insect cell expression system.The purified MICB molecules were coupled to Luminex beads.The linear range,specificity,precision and accuracy of the detection system were evaluated.A total of 500 samples of healthy individuals were tested by multiplex.Results MICB antibody multiplex flow detection assay was successfully established with a linear range from 3.1 to 100.0 μg/mL.The linearly dependent coefficient was 0.995.The rate of recovery was from 97.4% to 99.6% in recovery tests.The coefficients of variation（CV） of intra-assay were 3.4% and 2.9% in the concentrations of 30 μg/mL and 60 μg/mL,and the CV of inter-assay were 4.5% and 4.9% respectively.The assay was of high specificity without cross-reaction with MICA.MICB-specific antibodies were found in 4（0.8%） of 500 healthy adults.Conclusion The multiplex flow assay for MICB antibody showed high sensitivity,stability and accuracy and may be useful for the further study on the effect of MICB antibodies in organ transplantation.

关 键 词：MICB 流式荧光 等位基因多态性 抗体 

分 类 号：R446.6[医药卫生—诊断学]