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目的比较不同血清稀释滴度对儿童抗核抗体间接免疫荧光分析(ANA-IIF)结果的影响,探讨降低血清起始稀释度必要性。方法以间接免疫荧光法检测110例健康儿童系列稀释度血清标本抗核抗体(ANA),并与特异性ANA线性免疫(ANA-LIA)结果相比较;同时分析一组ANA-IIF阴性的临床患儿标本ANA-LIA结果。结果健康组标本随着稀释度从1∶80、1∶40、1∶20逐步减低,ANA-IIF阳性检出率有所上升,分别为7.3%、9.1%、10.9%,但差异无统计学意义(P>0.05),弱阳性分别为7.3%、15.5%、31.8%,差异有统计学意义(P<0.01)。110例健康体检儿童中8例标本检测出特异性ANA,阳性率为7.3%。8例IIF法稀释度1∶80阳性者中,特异性ANA阳性者2例;稀释度1∶40、1∶20新增的4例荧光ANA阳性标本中,有1例特异性ANA阳性。若视ANA-IIF(1∶80)或ANA-LIA任一阳性为ANA阳性,ANA阳性率从7.3%上升到12.7%。29例ANA-IIF(1∶80)为阴性的自身免疫肝病相关自身抗体检测患儿临床标本中,特异性ANA-LIA检测出5例阳性(17.2%)。结论降低儿童血清起始滴度并不能明显提高ANA-IIF阳性检出率,反而增加了非特异的弱阳性,因此,临床实验室不需改变儿童ANA常规标本稀释滴度,联合特异性ANA-LIA的检测有利于ANA的发现。
Objective To compare the effects of different serum dilutions on the results of indirect immunofluorescence assay (ANA-IIF) of antinuclear antibodies in children and to explore the necessity of reducing the initial dilution of serum. Methods ANA was detected by indirect immunofluorescence assay in 110 healthy children with serially diluted serum samples and compared with ANA-LIA. A group of ANA-IIF-negative clinical patients ANA-LIA results of pediatric specimens. Results The positive rate of ANA-IIF in healthy group decreased gradually from 1:80, 1:40 and 1:20 with the dilution of 7.3%, 9.1% and 10.9%, respectively, but the difference was not statistically significant Significance (P> 0.05), weak positive were 7.3%, 15.5%, 31.8%, the difference was statistically significant (P <0.01). In 110 cases of healthy children, 8 cases detected specific ANA, the positive rate was 7.3%. Among the 8 patients with 1:80 positive IIF dilution, 2 were positive for specific ANA, and 4 were positive for ANA in 4 of the 4 fluorescent ANA positive samples newly added at dilution 1:40 and 1:20. The ANA positive rate increased from 7.3% to 12.7% if either ANA-IIF (1:80) or ANA-LIA was positive for ANA. Of the 29 children with autoantigen-related autoantibodies tested negative for ANA-IIF (1:80), 5 were positive (17.2%) detected by specific ANA-LIA. Conclusion The reduction of serum initial titer in children does not significantly increase the positive rate of ANA-IIF, but increases the non-specific weak positive. Therefore, it is not necessary to change the dilution of routine ANA dilutions in clinical laboratories. Combined with specific ANA- The detection of LIA is in favor of the discovery of ANA.