[微阵列化学发光免疫分析法定量检测血清抗氨甲酰化蛋白自身抗体的建立及应用]
[Establishment and application of quantitative detection of serum anti-carbamylated protein autoantibodies by microarray chemiluminescence immunoassay].
作者信息
Yao Xiaoyang, Xue Miao, Gui Tiejun, Ma Chenyun, Jiang Xiudi, Yan Hongli
机构信息
Department of Laboratory Diagnosis, Changhai Hospital, Naval Military Medical University, Shanghai 200433; Department of Clinical Laboratory, Seventh People's Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai 200137, China.
Department of Laboratory Diagnosis, Changhai Hospital, Naval Military Medical University, Shanghai 200433, China.
出版信息
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2018 Oct;34(10):924-930.
Objective To develope a chemiluminescence immunoassay based on microarray protein chip technology for detecting the anti-carbamylated protein (CarP) antibody. We aimed to evaluate the detection performance of this method and to explore its preliminary clinical application value of this index in patients with rheumatoid arthritis (RA). Methods A quantitative detection method for anti-CarP antibody was established to evaluate the precision, minimum detection limit, linear range and specificity of the method. The 95th position of anti-CarP antibody level in serum of 120 healthy controls was defined as the cutoff value. The anti-CarP antibody level and positive rate in RA group and non-RA group were analyzed. And the correlation between anti-CarP antibody and disease activity index in RA group was analyzed. Results The precision of this method for detecting high-level sample and low-level sample was less than 15%; The linear range could reach (3.31~1448.18) AU/mL, and there was almost no cross-reaction between anti-CarP antibody and anti-CCP antibody. Compared with healthy control group, the level of anti-CarP antibody in RA group, anti-CCP antibody positive RA group and joint pain group was significantly higher, and that in undifferentiated connective tissue disease group was also higher. Compared with the 5% positive rate of anti-CarP antibody in healthy control group, the positive rate of RA patients was 28.21%, anti-CCP antibody positive RA patients was 32.2%, joint pain group was 38.89%, which were significantly higher. There was no statistical difference between other disease groups. The level of anti-CarP antibody was weakly correlated with level of rheumatoid factor (RF) and erythrocyte sedimentation rate (ESR) in RA patients, but it was moderately correlated with CRP and IgG level. Conclusion The protein chip chemiluminescence method for quantitative detection of anti-CarP antibody has good detection precision and wide linear range, and has good sensitivity and specificity. Anti-CarP antibody detection is valuable for RA diagnosis and disease activity evaluation.
目的 基于微阵列蛋白芯片技术开发一种化学发光免疫分析法用于检测抗瓜氨酸化蛋白（CarP）抗体。旨在评估该方法的检测性能，并探讨该指标在类风湿关节炎（RA）患者中的初步临床应用价值。方法 建立抗CarP抗体定量检测方法，评估该方法的精密度、最低检测限、线性范围和特异性。将120例健康对照者血清中抗CarP抗体水平的第95百分位数定义为临界值。分析RA组和非RA组抗CarP抗体水平及阳性率。并分析RA组抗CarP抗体与疾病活动指数的相关性。结果 该方法检测高、低水平样本的精密度均小于15%；线性范围可达（3.31~1448.18）AU/mL，抗CarP抗体与抗环瓜氨酸肽（CCP）抗体之间几乎无交叉反应。与健康对照组相比，RA组、抗CCP抗体阳性RA组和关节疼痛组抗CarP抗体水平显著升高，未分化结缔组织病组也升高。与健康对照组抗CarP抗体5%的阳性率相比，RA患者阳性率为28.21%，抗CCP抗体阳性RA患者为32.2%，关节疼痛组为38.89%，均显著升高。其他疾病组之间无统计学差异。RA患者中抗CarP抗体水平与类风湿因子（RF）和红细胞沉降率（ESR）水平呈弱相关，但与C反应蛋白（CRP）和IgG水平呈中度相关。结论 蛋白芯片化学发光法定量检测抗CarP抗体具有良好的检测精密度和较宽的线性范围，且具有较好的敏感性和特异性。抗CarP抗体检测对RA诊断及疾病活动评估具有重要价值。
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