人高迁移率族蛋白B1光激化学发光分析法的建立及应用  

Establishment and application of light initiated chemiluminescence assay of high mobility group box 1

作　　者：俞蕾[1] 肖华龙[1] 刘洁[2] 黄飚[3] 盛慧明[4] 张艺[3] 胡志刚[1] Yu Lei;Xiao Hualong;Liu Jie;Huang Biao;Sheng Huiming;Zhang Yi;Hu Zhigang(Clinical Laboratory,Wuxi People＇s Hospital,Nanfing Medical University,Wuxi 214023,China（Yu L,Xiao HL,Hu Z G;Respiration Medicine,Wuxi People＇s Hospital,Nanfing Medical University,Wuxi 214023,China（Liu J;Jiangsu Institute of Nuclear Medicine,Wuxi 214063,China（Huang B,Zhang Y;Clini-cal Laboratory,Tongren Hospital,Shanghai Jiao Tong University,Shanghai 200051,China（Sheng HM)

机构地区：[1]南京医科大学附属无锡市人民医院检验科,214023 [2]南京医科大学附属无锡市人民医院呼吸内科,214023 [3]江苏省原子医学研究所,无锡214063 [4]上海交通大学附属同仁医院检验科,200051

出　　处：《中华核医学与分子影像杂志》2018年第7期489-492,共4页Chinese Journal of Nuclear Medicine and Molecular Imaging

摘　　要：目的采用光激化学发光分析（LICA）技术建立快速定量检测人高迁移率族蛋白B1（HMGB1）的方法。方法采用2株配对的HMGB1单克隆抗体（简称单抗），1株单抗包被受体微球，另1株单抗先用生物素标记，再与链霉亲合素的供体微球共同组成人HMGB1的LICA检测方法，进而优化反应体系并对方法的各项性能指标进行评价。分别测定普通肺炎患者（35例）和重症肺炎患者（25例）血清HMGB1的浓度，并与健康体检者（35名）进行比较。结果建立的HMGB1 LICA检测方法的灵敏度为0.1 μg/L，线性测量范围为0.1～1 000 μg/L；批内和批间精密度分别为1.74%～2.92%和1.93%～3.73%，均低于5%；回收率范围为94.53%～106.37%，平均回收率为99.74%；与酶联免疫吸附测定方法有良好的相关性（r＝0.888 2）；与HMGB2和HMGB3重组抗原无明显交叉反应，特异性良好。普通肺炎患者和重症肺炎患者血清HMGB1的质量浓度分别为（6.76±3.13）和（19.69±9.04） μg/L，均高于健康体检者[（1.49±0.74） μg/L；t值：－5.447和－5.186，均P〈0.01]，普通和重症肺炎患者间差异亦有统计学意义（t＝－3.500，P〈0.01）。结论LICA法检测HMGB1灵敏度高、特异性强、结果可靠，且具有均相、快速、免清洗等特点，有良好的临床应用前景。ObjectiveTo establish a fast and quantitative light initiated chemiluminescent assay （LICA） method for high mobility group box1 （HMGB1） determination.MethodsTwo strains of paired HMGB1 monoclonal antibodies were used. One was used to coat receptor microspheres. The other was labeled with biotin first and then composed with chain mildew element of affinity donor microsphere to form LICA method for HMGB1. After optimizing the reaction system, the technical specifications of the method was evaluated. Serum HMGB1 levels of common pneumonia patients （CPP） and severe pneumonia patients （SPP） were measured and compared with that of health controls. Two-sample t test was used.ResultsThe sensitivity of LICA was 0.1 μg/L, with linear measurement ranging from 0.1 to 1 000 μg/L. The precisions of intra- and inter-analysis were 1.74%-2.92% and 1.93%-3.73% respectively, both were lower than 5%. The recovery rate was 99.74% （range： 94.53%-106.37%）. The correlation coefficient of LICA and enzyme-linked immunosorbent assay （ELISA） was 0.888 2. The LICA method had good specificity and no obvious cross reaction with HMGB2 and HMGB3. The serum HMGB1 level in CPP （n=35） and SPP （n=25） was significantly higher than that in health controls （n=35）： （6.76±3.13）, （19.69±9.04） vs （1.49±0.74） μg/L; t values： -5.447 and -5.186, both P〈0.01. The HMGB1 levels between CPP and SPP were also significantly different （t=-3.500, P〈0.01）.ConclusionsThe established LICA method of HMGB1 has high sensitivity and specificity with reliable results. This method is also homogeneous, fast and cleaning-free, thus has a good prospect in clinical application.

关 键 词：高迁移率族蛋白质类 化学发光测定法 肺炎 

分 类 号：R446.6[医药卫生—诊断学]