胶乳增强免疫比浊法检测抗环瓜氨酸肽抗体试剂盒的制备及性能评价  

Preparation and Performance Evaluation of Latex-enhanced Immunoturbidimetry Kit for Detection of Anti-cyclic Citrullinated Peptide Antibody

作　　者：金耀建[1] 黄信用 高素华[1] 叶均[1] 楼宏强[1] 张艳芳 孙义凯 JIN Yaojian;HUANG Xinyong;GAO Suhua;YE Jun;LOU Hongqiang;ZHANG Yanfang;SUN Yikai(Jinhua Polytechnic,Jinhua 321007,China;Qiangsheng Biotechnology Co.,Ltd.,Jinhua 321000,China)

机构地区：[1]金华职业技术学院,浙江金华321007 [2]金华市强盛生物科技有限公司,浙江金华321000

出　　处：《金华职业技术学院学报》2022年第3期64-70,共7页Journal of Jinhua Polytechnic

基　　金：浙江省基础公益研究计划项目(LGF20H200003)。

摘　　要：目的:胶乳增强免疫比浊法检测抗环瓜氨酸肽抗体试剂盒的制备及性能评价。方法:对反应体系中的主要成分定标,校准品溯源,质控品定值,参考区间确定,并对试剂准确度、精密度、稳定性、干扰物实验等相关性能指标进行评价,检测临床标本评价研制的试剂盒效能。结果:羧基微球粒径采用108 nm,羧基微球与环瓜氨酸肽抗原比例为3∶1,交联时间为60分钟,BSA封闭时间为60分钟,这时方法学灵敏度最佳。标准品溯源至北京九强生物技术股份有限公司试剂盒,两者线性回归y=1.010x-0.420(r=0.981)。质控品定值分别为20.0 U/mL±4.0 U/mL,50.0 U/mL±10.0 U/mL。参考范围95%可信区间为<35 U/mL。在2℃~8℃环境下,产品校准品贮存12个月和质控品开封16天后检测结果变化不显著。干扰物胆红素≤200 mg/L、甘油三酯≤3 g/L、血红蛋白≤2.5 g/L时,对三批试剂结果均无显著干扰。对照试剂和考核试剂检测临床标本,线性回归y=0.9607x-0.3451(r=0.9763),偏倚分析在允许误差范围内,说明考核试剂完全达到临床检测要求。结论:制备的胶乳增强免疫比浊法检测抗环瓜氨酸肽抗体试剂盒符合临床诊断要求。objective:To prepare and evaluate the performance of latex-enhanced immunoturbidimetry kit for detection of anti-cyclic citrullinated peptide antibody.Methods:The main components in the reaction system of the kit were calibrated.The traceability of calibrators,value of quality control products,and determination of reference interval in the kit were made.Further evaluations were made for its accuracy,precision,reagent stability,disruptor experiment and other related performance indexes.Clinical samples were also detected to evaluate the efficiency of the developed kit.Results:The particle size of carboxyl microsphere was 108 nm and the ratio of carboxyl microsphere to cyclic citrullinated peptide antigen was 3∶1.Both the crosslinking time and BSA blocking time were set to 60 minutes.The methodological sensitivity peaked with this experimental setup.The standard product was traced to the kit of Beijing Jiuqiang Biotechnology Co.,Ltd.,and linear regression of the two was y=1.010x-0.420(r=0.981).The values of the quality control products were 20.0 U/mL±4.0 U/mL and 50.0 U/mL±10.0 U/mL respectively.The 95%confidence interval of the reference range was<35 U/ml.In the environment of 2~8℃,the test results of product calibrators stored for 12 months and quality control products unsealed for 16 days did not change significantly.With bilirubin≤200 mg/L,triglyceride≤3 g/L and hemoglobin≤2.5 g/L as interferent,the results of three batches of reagents were not affected.The clinical samples were tested with control reagent and assessment reagent,and the linear regression was y=0.9607x-0.3451(r=0.9763).The bias analysis was within the allowable error range,indicating that the assessed reagent fully met the requirements of clinical testing.Conclusion:The latex-enhanced immunoturbidimetry kit for detection of anti-cyclic citrullinated peptide antibody meets the requirements of clinical diagnosis.

关 键 词：胶乳增强免疫比浊法 抗环瓜氨酸肽抗体 环瓜氨酸肽 类风湿关节炎 

分 类 号：R392-33[医药卫生—免疫学]