间接免疫荧光法与免疫印迹法检测抗核抗体的结果对比分析

Comparison of the Results of Indirect Immunofluorescence and Western blotting in the Detection of Anti-nuclear Antibodies

目的对比分析间接免疫荧光法检测抗核抗体(ANA)与免疫印迹法检测ANA谱的结果。方法选择2018年11月—2019年6月医院检验科接收的980份疑似自身免疫性疾病的血样本为实验对象,均采用间接免疫荧光法检测ANA和免疫印迹法检测ANA谱,对比分析两种方法的检测结果。结果①980份血样本中,间接免疫荧光法检测ANA阳性率26.84%,免疫印迹法检测阳性率23.16%,两组检出阳性率对比差异无统计学意义(χ^2=3.527,P=0.060)。但间接免疫荧光法诊断自身免疫性疾病的特异性高于免疫印迹法(χ^2=18.766,P=0.000),而免疫印迹法的敏感性却显著高于间接免疫荧光法(χ^2=17.084,P=0.000)。②在227例免疫印迹法检测的阳性样本中,有128例在间接免疫荧光法中呈斑点型,39例呈胞浆颗粒型,18例呈着丝点型,8例呈均质型,7例呈核仁型,4例呈核膜型,8例呈其他核型,另有15例间接免疫荧光法检测为阴性,而且免疫印迹法检测的阳性样本抗体类型不同其荧光核型也会有显著的差异。结论间接免疫荧光法检测ANA特异性较高,免疫印迹法检测ANA谱敏感性高,而且不同荧光核型抗核抗体与免疫印迹法检测的抗核抗体谱具有一定的相关性,但无绝对的规律可循,故在临床应用中两种检测方法不能相互替代,应联合应用以提高自身免疫性疾病诊断率。

Objective To compare the results of indirect immunofluorescence detection of anti-nuclear antibody(ANA)and immunoblotting detection of ANA spectrum.Methods From November 2018 to June 2019,980 blood samples suspected of autoimmune diseases received by the Hospital Laboratory Department were selected as the test subjects.Both indirect immunofluorescence method was used to detect ANA and immunoblotting method was used to detect ANA spectrum.The test results of the two methods were compared.Results 1.Among 980 blood samples,the positive rate of ANA detected by indirect immunofluorescence method was 26.84%,and the positive rate detected by immunoblotting method was 23.16%.There was no statistically significant difference between the positive rates of the two groups(χ^2=3.527,P=0.060).However,the specificity of indirect immunofluorescence for diagnosis of autoimmune diseases was higher than that of immunoblotting(χ^2=18.766,P=0.000),while the sensitivity of immunoblotting was significantly higher than that of indirect immunofluorescence(χ^2=17.084,P=0.000).2.Of the 227 positive samples detected by immunoblotting,128 were spotted in indirect immunofluorescence,39 were cytoplasmic,18 were centromere,8 were homogeneous,7 cases were nucleolar type,4 cases were nuclear membrane type,8 cases were other karyotypes,and another 15 cases were negative by indirect immunofluorescence method,and the antibody type detected by immunoblotting in positive was different.The fluorescent karyotype was also significant difference.Conclusion The indirect immunofluorescence method has high specificity for ANA detection,and the Western blot method has high sensitivity for detection of ANA spectrum.Moreover,anti-nuclear antibodies with different fluorescent karyotypes have a certain correlation with the anti-nuclear antibody spectrum detected by immunoblotting,but there is no absolute rules to be followed,so in clinical applications,the two detection methods cannot be replaced by each other,and should be used in combination to improve the diagnosis rate of autoimmune diseases.