竞争抑制法检测HBeAb的影响因素及对策

Influential factors in the detection of total anti-HBe antibody by competitive enzyme-linked imm unosorbentassay

目的研究酶联免疫吸附试验(ELISA)检测血清标本与辣根过氧化物酶(HRP)标记的乙型肝炎病毒(HBV)e抗体(抗-HBe-HRP),加入间隔时间对抗-HBe抗体检测结果的影响。方法选择HBV标志物检测结果为阴性的标本分为A、B、C 3组。A组加样后延长室温放置不同时间再加入抗-HBe-HRP;B组在加样的同时加入了抗-HBe-HRP延长室温放置不同时间;C组为A组方式测定的阴性转阳性标本再用ELISA试剂2和化学发光(CLIA)试剂重复测定,确认阴、阳性结果。结果A组加样方式对抗-HBe抗体检测结果影响明显,且加样和加入抗-HBe-HRP的间隔时间越长,标本的假阳性越多,B组加样方式对抗-HBe抗体检测结果基本无影响;C组ELISA试剂1和试剂2有73.1%的检测结果一致;25例ELISA阴性转阳性标本经过CLIA方法验证后仍为阴性。结论A组的不公平竞争在不同时间内可出现不同程度的抗-HBe抗体假阳性;B组加样方式可最大限度地减少抗-HBe抗体假阳性,保证了检验质量。

Objective To investigate the effect of interval between the addition to serum sample and anti-HBe-horseradish peroxidase（HRP）conjugate on the detection result of total ant i-HBe antibody. Methods Serum anti-HBe was detected by domestic enzyme linked immunosorbent assay f ELISA） kits and by Abbott chemiluminescent immunoassay（CLIA）system as aeontrol method. 72 pretested an ti-HBe negative samples were divided into three groups. GroupA： anti-HBe-HRP was added after the addition of serum sample and deposited at room temperature for different time（unfair competition）; Group B：the serum sample and anti-HBe HRP were added simutaneously and then deposited at room temperature for diferent time（fair competition） ;Group C：the positive samples in group A were detected by another domestic ELISA reagent according to the saIfle intervals and tested finally, by abbott CLIA method. Results The pattern of addition of serum sample and anti-HBe HRP affected the real results significantly. Furthermore, the longer intervals between the two additions led to more false pos itive results. Whereas,simultaneous addition of serum and anti HBe-HRP had no effects on the real resuits. In group C, the results of 73. 1 percentage＇of positive samples were consistent with group A. Twenty positive samples in group A showed negative reactions by abbott CLIA method. Conclusion The interval between the addition of serum sample and anti-HBe-HRP could lead to false positive re suit,which can be avoided maximumly with simultaneous addition.