软骨寡聚基质蛋白荧光免疫层析定量检测方法建立
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河南省科技攻关项目(182102310685)


Development of a fluorescence immunochromatography for quantitative detection of cartilage oligomeric matrix protein
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    摘要:

    本研究旨在建立一种可用于类风湿性关节炎(rheumatoid arthritis,RA)辅助诊断的人软骨寡聚基质蛋白(cartilage oligomeric matrix protein,COMP)荧光层析检测方法。采用双抗体夹心法原理制备免疫层析试纸条,并进行性能评价及方法学对比。通过对临床样本的检测得到受试者工作特征(receiver operating characteristic,ROC)曲线,计算试纸条灵敏度、特异性、阳性和阴性预测值。线性范围为0.39–50.00 ng/mL;批内批间变异系数均小于15%;试纸条37℃加速破坏20 d,荧光信号强度变化范围在15%以内;与类风湿因子(rheumatoid factor,RF)、抗环瓜氨酸肽(anti-cyclic citrullinated peptide,anti-CCP)抗体无交叉反应;与ELISA试剂盒平行检测48份临床血清样本,相关性良好。采用本研究制备的试纸条检测样本,COMP区分RA患者和健康人的cut-off值为22.55 ng/mL (灵敏度为0.821,特异度为0.842,阳性预测值为0.741,阴性预测值为0.895)。同时使用ELISA试剂盒进行相同样本检测,两种检测方法灵敏度、特异度均能达到80%以上。这种可定量检测COMP的荧光层析试纸条具有快速、简便、灵敏等优点,可为RA患者提供快速辅助诊断。

    Abstract:

    The purpose of this study was to develop a fluorescence chromatography method for the detection of cartilage oligomeric matrix protein (COMP) in the auxiliary diagnosis of rheumatoid arthritis (RA). The principle of double antibody sandwich method was used to prepare immunochromatographic test strips, and the performance evaluation and methodological comparison were carried out. Through the detection of clinical samples, a receiver operating characteristic (ROC) curve was obtained, and the sensitivity, specificity, positive and negative predictive values of the test strip were calculated. The linear range was 0.39-50.00 ng/mL. The coefficients of variation inter and intra batches were less than 15%. The test strip was stable at 37℃ for 20 days, and the variation range of fluorescence signal intensity was within 15%. There was no cross reaction with rheumatoid factor (RF) and anti-cyclic citrulline peptide (anti-CCP) antibody. Forty-eight clinical serum samples were detected in parallel with ELISA kit, and the correlation was good. The test strip prepared in this study was used to detect the sample, the cut-off value of COMP between RA patients and healthy people was 22.55 ng/mL (sensitivity 0.821, specificity 0.842, positive predictive value 0.741, negative predictive value 0.895). At the same time, the same sample was tested with ELISA kit, the sensitivity and specificity of the two methods reached more than 80%. A quantitative COMP fluorescence chromatography test strip was developed, which has the advantages of celerity, simplicity and sensitivity, and may provide rapid auxiliary diagnosis for RA patients.

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张永生,王云龙,王继创,李玉林,王晓军,王旭东. 软骨寡聚基质蛋白荧光免疫层析定量检测方法建立[J]. 生物工程学报, 2022, 38(6): 2332-2341

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  • 收稿日期:2021-09-26
  • 在线发布日期: 2022-06-28
  • 出版日期: 2022-06-25
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