摘要
目的了解类风湿关节炎(RA),其他自身免疫病和健康对照组血清抗Ⅱ型胶原短肽,即CⅡ(260-272)抗体水平,探讨其在RA疾病诊断和病情监测中的价值。方法将合成的CⅡ(260-272)短肽与牛血清白蛋白(BSA)耦联得CⅡ(260-272)-BSA复合物,用其作为抗原包被建立抗CⅡ(260-272)抗体的ELISA测定方法,经方法学考核后,对临床病例标本检测结果进行评价。结果建立的抗CⅡ(260-272)抗体的ELISA法实验结果具有较好的稳定性和特异性,其批内和批间变异系数分别为8.4%和9.6%,特异阻断抑制率最高达76.3%。临床检测结果发现抗CⅡ(260-272)抗体在RA中的阳性率为75.0%(30/40),显著高于其他自身免疫病和健康对照组(P<0.01)。以测定吸光度(A)0.213为临界值时,抗CⅡ260-272抗体对RA诊断特异性91.0%,阳性预测值63.8%,阴性预测值94.5%,阳性似然比8.33,阴性似然比0.275,Youden指数0.66。ROC曲线分析结果显示该抗体对RA诊断准确度为中等偏上水平(AUCROC值=0.890)。未发现抗CⅡ(260-272)抗体与类风湿因子(RF)水平相关(在13例RF阴性RA患者中,该抗体阳性10例,阳性率76.9%),但抗CⅡ(260-272)抗体与抗环瓜氨酸肽(CCP)抗体显著相关(P<0.01),且该抗体水平随患者临床症状的改善有逐渐降低趋势。结论成功建立抗CⅡ(260-272)抗体ELISA测定方法,用其检测抗CⅡ(260-272)抗体水平可作为RA疾病诊断和病情监测的一种新的参考指标。
Objective To understand serum levels of anti-type Ⅱ collagen peptide,namely CⅡ(260-272) antibody in rheumatoid arthritis(RA),other autoimmune diseases and healthy control,and explore the value of this antibody in diagnosis and monitoring of RA disease.Methods Synthetic peptide of CⅡ(260-272) was coupled to bovine serum albumin(CⅡ(260-272)-BSA),using this conjugate as coated antigen,ELISA method was established for anti-CⅡ(260-272) antibody.After its methodology assessed,results of clinical cases tested by this method were evaluated.Results The establishment of ELISA method for anti-CⅡ(260-272) antibody has the better results of stability and specificity,and its intra-and inter-assay coefficients of variation were 8.4% and 9.6% respectively.The specific inhibitory rate of this method was up to 76.3%.Clinical test results found that the positive rate of anti-CⅡ(260-272) antibody was 75.0%(30/40) in RA,this result was significantly higher than that of other autoimmune diseases and healthy control group(P〈0.01).When absorbance of 0.213 was considered cut-off value,the anti-CⅡ(260-272) antibody diagnostic specificity,positive/negative predictive value,positive/negative likelihood ratio and Youden's index was 91.0%,63.8%/94.5%,8.33/0.275 and 0.66 in RA patients,respectively.ROC curve analysis revealed that this antibody on the accuracy of RA diagnosis was at middle level(AUCROC value= 0.890).No correlations was found between level of anti-CⅡ(260-272) antibody and rheumatoid factor(RF)(in 13 cases of RF-negative RA patients,10 cases of anti-CⅡ(260-272) antibody were positive,positive rate was 76.9%),but significantly correlated with anti-cyclic citrullinated peptide(CCP) antibody(P〈0.01),furthermore,serum levels of anti-CⅡ(260-272) antibody decreased with the improvement of clinical symptoms of RA cases.Conclusion ELISA method for anti-CⅡ(260-272) antibody was successful established,serum levels of this antibody to be tested could be used as a new reference for diagnosis and condition monitoring of RA disease.
出处
《中国实验诊断学》
北大核心
2009年第12期1714-1718,共5页
Chinese Journal of Laboratory Diagnosis
基金
中国博士后基金特别资助项目(200801375)
关键词
Ⅱ型胶原
短肽
类风湿关节炎
酶联免疫吸附试验
type Ⅱ collagen
peptide
rheumatoid arthritis
enzyme-linked immunosorbent assay