间接免疫荧光法与ELISA检测抗核抗体、抗双链DNA抗体的比对分析  

Comparison of indirect immunofluorescence assay and ELISA for detecting antinuclear antibodies and anti-double-stranded DNA antibodies

作　　者：秦雪[1] 陶瑕[2] 陈志坚[1] 蒋杰球[1] 徐明辉[3] 李若林[3] 李泰阶[3] 林发全[1] 李山[1] 

机构地区：[1]广西医科大学第一附属医院临床医学实验部,广西南宁530021 [2]广西医科大学2003级医学检验本科,广西南宁530021 [3]广西医科大学研究生学院,广西南宁530021

出　　处：《南方医科大学学报》2009年第3期472-475,共4页Journal of Southern Medical University

基　　金：广西卫生厅科研课题(z2008158)

摘　　要：目的探讨间接免疫荧光法(IIFA)和酶联免疫吸附试验(ELISA)检测抗核抗体(ANA)和抗双链DNA(ds-DNA)抗体的异同。方法抽取确诊或疑为自身免疫性疾病患者血清125例,同时采用IIFA法和ELISA法检测ANA82例,抗ds-DNA抗体检测57例,其中14例同时检测ANA和抗ds-DNA抗体。对两种方法检测结果不符者采用免疫印迹法复查抗可溶性核抗原(ENA)。结果82例ANA阳性率IIFA法与ELISA法分别为87.8%和73.17%,差别有统计学意义(P<0.01);57例血清中抗ds-DNA抗体检测阳性率IIFA法与ELISA法分别为77.19%和71.93%,差别无统计学意义(P>0.05)。采用不同cutoff值时两种方法符合率比较,ANA、抗ds-DNA抗体检测结果差别均有统计学意义(P<0.01)。两法对一些稀有核型ANA检测符合率较低,而抗ds-DNA抗体的符合率在不同核型上没有差异。ANA检测以1:100为cutoff滴度,抗ds-DNA抗体以弱阳性为cutoff值时,两种方法可获得较高的符合率,但在检测滴度1:100ANA、弱阳性抗ds-DNA抗体时两种方法存在明显的不一致。结论IIFA法检测总ANA、抗ds-DNA抗体的敏感性均高于ELISA法,IIFA法作为ANA的筛查方法,阳性标本再进行ELISA法检测,可在获得荧光核型信息同时观察抗体滴度的变化情况,并且更快捷经济。两种或多种不同方法学原理的实验互相佐证,有助于减少实验误差。Objective To compare indirect immunofluorescence assay （IIFA） and enzyme-linked immunosorbent assay （ELISA） for detecting antinuclear antibodies （ANA） and anti-double-stranded DNA antibodies （anti-dsDNA）. Methods A total of 125 serum samples were obtained from patients with established or suspected autoimmune disease, and 82 samples were used for ANA detection and 57 for anti-dsDNA detection using both IIFA and ELISA. Fourteen samples were examined for both ANA and anti-dsDNA. In cases where discrepancy occurred in the results by the two methods, extractable nuclear antigens were detected using immunoblotting. Results The positivity rate of ANA detected by IIFA and ELISA was significantly different （87.8% and 73.17%, respectively, P〈0.01）, but the positivity rate of anti-dsDNA was similar between IIFA and ELISA （77.19% and 71.93%, respectively, P〉0.05）. The percent agreement between the two testing methods with different cutoff values of ANA and anti-dsDNA showed significant differences （P〈0.01）, and for some uncommon patterns, the percent agreement of the two methods was lowered in ANA detection but remained unchanged for anti-dsDNA with different ANA patterns. High percent agreements of the two methods were obtained with the cutoff ANA titer of 1：100 and the cutoff anti-dsDNA value of weak positivity, but they demonstrated a significant difference in testing low-titer ANA and anti-dsDNA. Conclusion IIFA is more sensitive than ELISA in detecting the total ANA and anti-dsDNA. ELISA prescreening combined with IIFA can obtain the information of the nuclear pattern and allow the observation of the titer alterations. The combination of two or more testing methods can greatly enhance the accuracy of the results.

关 键 词：抗核抗体 抗双链DNA抗体 酶联免疫吸附试验 间接免疫荧光法 

分 类 号：R446.6[医药卫生—诊断学]