预包被抗体结合抗原包被方式酶联免疫吸附试验共系统检测乙型肝炎病毒e抗原结果分析

Analysis on the detection of sample HBeAg by enzyme-linked immunosorbent assay in a common measurement system with anti-HBe using its ELISA reagent kits of pre-coating polyclonal anti-HBe binding recombinant HBeAg

目的探讨用竞争抑制法检测预包被抗体结合抗原包被方式[抗乙型肝炎病毒(HBe)EIA]商品试剂盒共系统检测乙型肝炎病毒e抗原(HBeAg)的可行性及结果判定方法。方法用竞争抑制法检测抗HBe酶联免疫吸附试验(ELISA)试剂盒共系统检测1000份随机血清(浆)标本抗HBe与HBeAg,并以常规ELISA双抗体夹心法检测HBeAg试剂盒检测HBeAg作平行对照。结果ELISA共系统检测HBeAg,其阳性检测孔显色较抗HBe阴性对照孔(或HBeAg阴性参比对照孔)明显加深,易于目测;其比色判定临界值(COV)易于设定(或使用HBeAg临界值血清),与常规ELISA检测HBeAg比色判定差异无统计学意义(配对计数资料比较的χ2检验:相关检验均为P<0.005,υ=1,a=0.05;优劣检验依次为HBeAg临界值血清①P>0.9000,υ=1,a=0.05、中和(竞争)抑制法检测抗HBeEIA商品试剂盒中抗HBe阴性对照;②0.100>P>0.050,υ=1,a=0.05、ELISA共系统检测HBeAgCOV;③0.100>P>0.050,υ=1,a=0.05,但χ21<χ22<χ23)。结论用竞争抑制法检测抗HBeELISA商品试剂盒能够检测HBeAg,在有优质现代酶标仪的实验室,可以省去常规ELISA双抗体夹心法检测HBeAg试剂盒,实用价廉,值得推广。

Objective To investigate the practicality and how to deal with the results of detecting sample HBeAg by ELISA in a common measurement system with Anti-HBe using its enzyme-linked immunosorbent assay （ELISA）precoating polyclonal Anti-HBe binding recombinant （HBeAg） reagent kits of a competitive inhibition method.Methods HBeAg in 1 000 specimens random serum（or plasma）samples was measured by ELISA in a common measurement system with Anti-HBe using its ELISA reagent kits of a competitive inhibition method and the results were also detected by a routine ELISA of HBeAg using its EIA reagent kits of a double？antibody sandwich method as a control.Results The results of detecting sample HBeAg in the common mearurement system are easy to be differentiated with naked eyes,the colour of positive hole was obviously heavier than negative control of Anti-HBe （or negative reference control of HBeAg）;its cut off value （COV）was similar to a routine ELISA of HBeAg and easier to be set up （or HBeAg serum of COV was used）and the HBeAg quanlitative assay of OD measurements was no significant defference between ELISA in a common measurement system and a routine ELISA of HBeAg（χ^2-test of enumeration data with paried design：correlation test were all P〈0.005,υ=1,a=0.05;difference test is successively HBeAg serum of COV ① P〈0.900,υ=1,a=0.05;Anti-HBe negative control of its EIA reagent kits of a nuetralizing and competitive inhibition method ②0.100P〈0.050,υ=1,a= 0.05;COV of HBeAg EIA in a common measurement system ③ 0.100P〈0.050,υ=1,a=0.05,however χ21χ22χ23）.Conclusion The detection of sample HBeAg by ELISA in a common measurement system with Anti-HBe using its ELISA reagent kits of a competitive inhibition method is practicable,if a modern and high quality analyzer of ELISA were installed in one＇s own clinical laboratory,a routine ELISA （a double-antibody sandwich method）reagent kits of HBeAg should be ommitted.It is of practicable and cheap and is worthy of popularizing.