Qin Xue, Tao Xia, Chen Zhi-Jian, Jiang Jie-Qiu, Xu Ming-Hui, Li Ruo-Lin, Li Tai-Jie, Lin Fa-Quan, Li Shan
Department of Clinical Laboratory, First Affiliated Hospital, Guangxi Medical University, Nanning, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2009 Mar;29(3):472-5.
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).
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.
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.
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.
比较间接免疫荧光法(IIFA)和酶联免疫吸附测定法(ELISA)检测抗核抗体(ANA)和抗双链DNA抗体(抗dsDNA)的效果。
从确诊或疑似自身免疫性疾病患者中获取125份血清样本,其中82份用于ANA检测,57份用于抗dsDNA检测,同时采用IIFA和ELISA两种方法。14份样本同时检测ANA和抗dsDNA。若两种方法结果出现差异,则采用免疫印迹法检测可提取核抗原。
IIFA和ELISA检测ANA的阳性率差异有统计学意义(分别为87.8%和73.17%,P<0.01),但IIFA和ELISA检测抗dsDNA的阳性率相似(分别为77.19%和71.93%,P>0.05)。两种检测方法在不同ANA和抗dsDNA临界值下的一致性百分比差异有统计学意义(P<0.01),对于一些不常见模式,两种方法在ANA检测中的一致性百分比降低,但在不同ANA模式下抗dsDNA检测的一致性百分比保持不变。当ANA临界滴度为1:100和抗dsDNA弱阳性临界值时,两种方法的一致性百分比很高,但在检测低滴度ANA和抗dsDNA时存在显著差异。
IIFA在检测总ANA和抗dsDNA方面比ELISA更敏感。ELISA预筛查结合IIFA可获得核型信息并观察滴度变化。两种或更多检测方法联合使用可大大提高结果的准确性。