单纯疱疹病毒IgM抗体(1+2型)时间分辨免疫荧光分析法的建立  

Time-resolved fluoroimmunoassay for herpes simplex virus type 1 and type 2 IgM antibodies

作　　者：杨运森[1,2] 周健伟 董志宁 刘天才[1] 吴英松[1] 

机构地区：[1]南方医科大学检验与生物技术学院,广东广州510515 [2]中国人民解放军第一八一医院医务处,广西桂林541002 [3]广州市达瑞生物技术股份有限公司,广东广州510130

出　　处：《热带医学杂志》2016年第12期1483-1487,共5页Journal of Tropical Medicine

基　　金：国家科技部重大项目(2013ZX10004-803);国家自然科学基金(81271931)

摘　　要：目的利用时间分辨免疫荧光分析法研制单纯疱疹病毒IgM抗体(1+2型)检测试剂盒。方法基于"捕获法"原理,在96孔酶标板上包被抗人IgM单克隆抗体,铕标记特异性单纯疱疹病毒1型和2型单克隆抗体作为示踪物,建立单纯疱疹病毒IgM抗体(1+2型)检测试剂,并对自研试剂盒的各项性能进行评价,与同类试剂盒进行方法学比对试验。结果自研试剂盒的灵敏度与ELISA比较有较大的提高,分析内变异系数≤4.29%,分析间变异系数≤6.15%;血清盘检测结果与该血清盘说明书提供的Gull IFA试剂的测试结果一致;与进口酶免法试剂同时检测293份临床样本,自制试剂盒的临床敏感性达100%,临床特异性达98.88%,Kappa值为0.940(P<0.001)。结论自研试剂盒灵敏度高、特异性强、精密度好,与商业化的同类产品检测结果一致性好,能满足临床应用的需要。Objective A novel time-resolved fluoroimmunoassay （TRFIA） methodology was developed for the determination of HSV IgM in human serum. Methods The assay was performed in 96-well microtiter plates coated with anti-IgM immunoglobulin. Eu^3＋-labeled HSV-1 and HSV-2 monoclonal antibodies were used as tracers. The performance of self-test reagents was evaluated, compared with the similar kit. Results The assay sensitivity was higher than ELISA, the intra- and inter assay coefficients of variation were less than 4.29% and 6.15%, respectively. Serum panel tests were performed using TRFIA and the results were satisfactory. 293 clinical samples were tested simultaneously with ELISAs. TRFIA was highly sensitive （100%） and specific （98.88%）, with a kappa value of 0.940. This result was in strong agreement with commercial ELISAs. Conclusion An effective, sensitive and specific detection of HSV type 1 and type 2 IgM TRFIA was successfully developed for diagnosis and clinical application.

关 键 词：单纯疱疹病毒 IGM抗体 时间分辨免疫荧光分析法 

分 类 号：R446.62[医药卫生—诊断学]