基于纳米磁微粒化学发光分析法的腐食酪螨过敏原特异性IgE抗体检测方法的建立

Establishment of specific IgE antibody detection method for Tyrophagus putrescentiae allergen based on nano-magnetic particle chemiluminescence analysis

目的建立用于腐食酪螨过敏原特异性IgE抗体检测的纳米磁微粒化学发光法。方法依据化学发光常规操作步骤,设置条件建立腐食酪螨过敏原特异性IgE检测的化学发光反应体系和合适的免疫反应条件,并以美国Phadia方法为标准,评估所建立化学发光方法的检验性能。应用SPSS21.0软件对两种方法的阳性率x2进行检验,以Pha出a为标准,评价化学发光方法的检测性能。结果化学发光反应体系中,发光底物A液中鲁米诺浓度为0.4mg/ml、发光底物B液中过氧化氢脲浓度为0.2mg/ml,此发光反应体系对辣根过氧化物酶(horseradish peroxidase,HRP)的敏感性很髙,可检测到H RP的最小量为0.01mg/m l,在免疫反应中,加人底物A液和B液后室温避光反应5min检测,在5〜30 min内检测数值均有效,37 t:为最佳发光反应温度;选择0.35~H X H U/m l范围作为标准曲线;以Phadia方法为标准,本研究建立的化学发光法敏感度为80.95%、特异度为100%、漏诊率为19%,用于检査腐食酪螨有显著意义。结论本研究成功建立纳米磁微粒化学发光法检测腐食酪蜗过敏原特异性IgE抗体。

Objective To establish a nanometer magnetic particle chemiluminescence methodfor the detection of specific IgE antibodies to Tyrophagus putrefaciens allergen.Methods According to the routine operation steps of chemiluminescence,chemiluminescence reaction system and suitable immune reaction conditions for detection of specific IgE of Tyrophagus putrescentiae allergen are established.Considering the performance of the chemiluminescence method established in this paper,the American Somerfly Phadia m ethod wasused as the gold standard to evaluate the test performance of the established chemiluminescence method.Results In thechemiluminescence reaction system,Luminol concentration in luminescent substrate A solution is 0.4 mg/ml,Urea hydrogen peroxidein luminescent substrate B solutionis 0.2 m g/ml.The sensitivity of this luminescent reaction system to horseradish peroxidase(H R P)is very high,the m i n i m u m detectable HRP is 0.01 n g/m l.In the immune response,the roo m temperature light-shielding reaction was detected for 5 min after the addition of substrates A and B,and the detection values were effective within 5-30 m i n.37^is optimum luminescence reaction temperature.Choose the range of 0.35-100 IU/ml as the standard curve.According to the results of 120 clinical trials,using Phadia(immunofluorescence method)as the gold standard,the chemiluminescence method established in this study has significant significance for the detection of Tyrophagus putrescentiae allergen.Conclusion A nanometer magnetic particle chemiluminescence method was successfully established for the detection of specific IgE antibodies to Tyrophagus putrescentiae allergen.