抗ANXA1自身抗体ELISA检测方法的建立及应用  

Establishment and application of ELISA methodology for the detection of anti ANXA1 autoantibody

作　　者：黄译乐 胡晓强 叶敏玲 徐建华 李琳[1] 吴祖培[2] 陈宇宁[3] 何金勇 梁玉瑶 HUANG Yi-le;HU Xiao-qiang;YE Min-ling;XU Jian-hua;LI Lin;WU Zu-pei;CHEN Yu-ning;HE Jin-yong;LIANG Yu-yao(Department of Clinical Laboratory,Guangzhou University of Traditional Chinese Medicine,Foshan 523800,China;Department of Oncology,Guangzhou University of Traditional Chinese Medicine,Foshan 523800,China;Department of Breast Surgery,Shunde Hospital,Guangzhou University of Traditional Chinese Medicine,Foshan 523800,China)

机构地区：[1]广州中医药大学顺德医院检验科,广东佛山523800 [2]广州中医药大学顺德医院肿瘤科,广东佛山523800 [3]广州中医药大学顺德医院乳腺外科,广东佛山523800

出　　处：《生物技术》2022年第5期565-568,575,共5页Biotechnology

基　　金：广东省自然科学基金面上项目(2019A1515011960);佛山市医学类科技攻关项目(2020001004538);佛山市卫生健康局医学科研课题(20210369);佛山市新型冠状病毒感染的肺炎应急科技攻关专项(2020001000431)。

摘　　要：[目的]建立检测抗膜联蛋白A1(Annexin A1,ANXA1)自身抗体的ELISA方法,初步探索其临床应用。[方法]设计ANXA1特异性抗原肽并人工合成,用抗原肽制备可检测ANXA1抗体的ELISA板,通过回归方程的建立、精密度的测定和抗干扰能力的验证,建立检测ANXA1抗体的ELISA方法。用该法检测临床乳腺癌及健康对照患者血清中的抗ANXA1自身抗体。[结果]该方法在抗体浓度0.025~0.400μg/mL范围内线性良好,R2=0.994,连续5次批内和批间变异系数%均小于10%,对乳糜标本和溶血标本的检测相对误差均小于10%,用于乳腺癌患者抗ANXA1自身抗体的诊断上,实验组抗ANXA1自身抗体表达显著高于对照组,差异具有统计学意义(P<0.05)。[结论]建立了抗ANXA1自身抗体检测的ELISA方法,线性范围0.025~0.400μg/mL,变异系数小于10%,对乳糜标本和溶血标本检测的相对误差小于10%,应用于临床初步显示乳腺癌患者血清中抗ANXA1自身抗体高表达。[Objective]To establish the ELISA method for the detection of anti Annexin A1(ANXA1)autoantibody and explore its clinical application.[Method]The ANXA1 specific antigen peptide sequence,which was designed online and synthesized artificially,was coated on the ELISA plate for ANXA1 autoantibody detection.A stable and reliable ELISA method for ANXA1 autoantibody detection was established through the building of regression equation,the determination of precision and the verification of anti-interference ability.[Result]The method was stable in the linear concentration range of 0.025-0.400μg/mL,R2=0.0944.The coefficient of variation(CV%)of 5 consecutive intra-assay and inter-assay batches was less than 10%.The relative error of detection of chylous and hemolytic samples was less than 10%.The expression of ANXA1 autoantibody in patients with breast cancer was significantly higher than that in the control group.The difference was statistically significant(P<0.05).[Conclusion]The ELISA methodology(linear range:0.025-0.400μg/mL,the coefficient of variation<10%,and the relative error<10%while the detection was disturbed by chylous and hemolytic specimens)for the detection of ANXA1 autoantibody was established.High expression of ANXA1 autoantibody was shown in serum of patients with breast cancer.

关 键 词：ANXA1 自身抗体 ELISA 乳腺癌 

分 类 号：R392.11[医药卫生—免疫学]