电化学发光免疫分析法定量测定HBsAg最适稀释度的研究

STUDY ON THE OPTIMAL DILUTION OF SERUM FOR QUANTITATIVE MEASURING LEVEL OF HEP ATITIS B SURFACE ANTIGEN BY ELECTROCHEMILUMINESCENCE IMMUNOASSAY

目的 :探讨电化学发光免疫分析法 (ECLIA)定量测定HBsAg的最适稀释度。方法 :采用正常混合血清倍比稀释HBsAg(+) /HBeAg(+) /HBcAb(+)、HBsAg(+) /HBeAb(+) /HBcAb(+)和HBsAg(+) /HBcAb(+)三种模式血清 ,应用ECLIA夹心法定量测定其HBsAg的含量。结果 :ECLIA法定量测定HB sAg阳性的三种血清学模式血清稀释度不同 ,HBsAg(+) /HBeAg(+) /HBcAb(+)模式存在HD HOOK效应 ,其最适稀释度为 1 :32 ;HBsAg(+) /HBeAb(+) /HBcAb(+)和HBsAg(+) /HBcAb(+)模式不需进行稀释 ,原倍血清HBsAg测定值最高。结论 :利用ECLIA法定量测定HBsAg(+) /HBeAg(+) /HBcAb(+)模式血清中HBsAg时 ,应对其血清进行预稀释 ,同时测定原倍和 1 :32稀释血清HBsAg含量 ,根据测定情况计算其结果 ,而HBsAg(+) /HBeAb(+) /HBcAb(+)和HBsAg(+) /HBcAb(+)模式 ,其血清HBsAg测定一般不需进行预稀释。

Objectives: To study the optimal dilution of serum for quantitative measuring of level of hepatitis B surface antigen (HBsAg) by electrochemiluminescence immunoassay (ECLIA). Methods: The sera from the three serological patterns [HBsAg(+)/HBeAg(+)/HBcAb(+), HBsAg(+)/HBeAb(+)/HBcAb(+) and HBsAg(+)/HBcAb(+)]were collected and diluted to a serial of different concertration with normal negtive mixed sera. The level of HBsAg was dectected by ECLIA separatedly. Results: The optimal dilution of serum for HBsAg quantitative measuring in serological pattern of HBsAg(+)/HBeAg(+)/HBcAb(+) is at 32 folds because of high does(HD)-hook effect, while HBsAg level of non-diluted specimen is higher than that of diluted ones in two other serological patterns. Conclusions: In ECLIA, high does(HD)-hook effect results in less reactivity and false-negative results.In order to avoid the experimental misdiagnosis, it is necessary to predilute the serum specimen when the HBsAg concentrations exceed the measuring range of the ECLIA method. The results indicated that the specimen should be prediluted to 32 folds when measure the serum of HBsAg level in HBsAg(+)/HBeAg(+)/HBcAb(+) serological pattern, while there is no need to measuring of HBsAg level in two other serological patterns.