曲霉半乳甘露聚糖磁微粒化学发光法检测体系的建立及评价  

Establishment and evaluation of aspergillus galactomannan magnetic particle chemiluminescence detection system

作　　者：马梦男 高欣欣 刘春龙 张舟 王梦凡[1] MA Mengnan;GAO Xinxin;LIU Chunlong;ZHANG Zhou;WANG Mengfan(School of Chemical Engineering and Technology,Tianjin University,Tianjin 300072,China)

机构地区：[1]天津大学化工学院,天津300072 [2]“侵袭性真菌病机制研究与精准诊断”北京市重点实验室丹娜生物分中心,天津300467

出　　处：《中国免疫学杂志》2022年第21期2623-2627,2633,共6页Chinese Journal of Immunology

基　　金：天津市科技计划项目(15ZXQXSY00050)。

摘　　要：目的:制备抗曲霉半乳甘露聚糖(GM)的多克隆抗体,并基于所获抗体建立并评价曲霉半乳甘露聚糖磁微粒化学发光(CLIA)检测体系。方法:高纯度GM抗原作为免疫原制备高效价、交叉反应弱的多克隆抗体。以磁微粒偶联的链霉亲和素,生物素标志的抗GM抗体,待测血清样本和吖啶磺酰胺标志的抗GM抗体建立GM磁微粒化学发光夹心法检测体系,并对其进行条件优化和性能评估。结果:检测回收率(100±15)%;分析内精密度、分析间精密度均<10%;分析灵敏度LoB、LoD分别为0.05、0.08 ng/ml;线性区间为0.15~50.00 ng/ml;Kappa系数0.88>0.75,与市售伯乐试剂盒具有良好的一致性。结论:本研究建立的GM磁微粒化学发光定性检测方法性能良好,具有潜在的市场转化应用前景,为侵袭性曲霉病早期临床诊断提供有益技术借鉴。Objective:To prepare polyclonal antibodies against aspergillus galactomannan(GM),and to establish and evaluate the Aspergillus galactomannan magnetic particle chemiluminescence(CLIA)detection system based on the obtained antibodies.Methods:High-purity aspergillus GM antigen was used as immunogen to prepare high-titer,weakly cross-reactive polyclonal antibody.Streptavidin coupled with magnetic particles,biotin-labeled anti-aspergillus GM antibody,the serum sample to be tested and the antiaspergillus GM antibody labeled with acridine sulfonamide successfully established an aspergillus GM magnetic particle chemiluminescence sandwich detection system,and optimized its conditions and performance evaluation.Results:The detection recovery rate was(100±15)%.The precision within and between analyses were less than 10%.The analytical sensitivity of LoB and LoD were 0.05,0.08 ng/ml.The linear interval between 0.15~50.00 ng/ml.Kappa coefficient 0.88>0.75,which has good consistency with Bio-Rad kit.Conclusion:The aspergillus GM magnetic particle chemiluminescence qualitative detection method established in this study has good performance and has potential market transformation application prospects,which can provide a useful technical reference for early clinical diagnosis of invasive aspergillosis.

关 键 词：半乳甘露聚糖 多克隆抗体 磁微粒化学发光法 侵袭性曲霉病 性能评估 

分 类 号：R446.6[医药卫生—诊断学]