抗rSjCTPI单克隆抗体检测血吸虫循环抗原的研究

STUDY ON DETECTION OF CIRCULATING SCHISTOSOME ANTIGEN WITH ANTI-rSjCTPI MONOCLONAL ANTIBODY

为探讨抗重组血吸虫磷酸丙糖异构酶 (r Sj CTPI)的单克隆抗体检测血吸虫循环酶抗原在血吸虫病免疫诊断和疗效考核中的价值。制备日本血吸虫 r Sj CTPI表达蛋白 ,以此重组蛋白免疫小鼠 ,筛选出高效价的抗 r Sj CTPI单抗 ,建立以纯化的抗血吸虫可溶性虫卵抗原 (SEA)的多克隆抗体 (Ig G)为捕捉系统 ,以酶标的抗 r Sj CTPI单抗为检测系统的酶联免疫吸附试验 (P- M- EL ISA)法检测血吸虫病人等血清。并对该法的敏感性、特异性、交叉反应以及疗效考核与常规的 SEA- EL ISA比较。结果获得了一株抗 r Sj CTPI的单克隆抗体 ,其抗体同型为 Ig G1。P- M- EL ISA检测急性、慢性血吸虫病人、健康人以及华枝睾吸虫病人、肺吸虫病人血清的阳性率分别为 10 0 %、91.7%、 0、0和0。而 SEA- EL TSA为 10 0 %、98.3%、6 .6 9%、10 %和 2 0 %。P- M- EL ISA法对 30例同批血吸虫病人治疗前的阳性率和治愈后 4个月的阴转率分别为 10 0 %和 30 % ;而 SEA- EL TSA分别为 10 0 %和 10 %。结果表明 ,应用抗 r Sj CT-PI单抗检测血吸虫病人循环抗原的 P- M- EL ISA法具有较高的敏感性和特异性 ,并有一定的疗效考核价值。

To explore the value on immuno diagnosis and evaluation of chemotherapy efficacy of schistosomiasis with anti rSjCTPI monoclonal antibody. Preparing the expression protein of rSjCTPI, and immunizing mice with the protein to prepare the anti rSjCTPI monoclonal antibody. Establishing P M ELISA with anti SEA polyclonal antibody IgG to be as the capture antibody, and anti rSjCTPI monoclonal antibody conjugated HRP to be as detecting antibody. Testing the sensitivity, specificity, cross reaction and evaluation of chemotherapy efficacy of P M ELISA, and compared with SEA ELISA. The results showed that one of high efficient monoclonal antibody was obtained, its isotope was IgG1. The positive rates of P M ELISA for detecting acute, chronic schistosomiasis, healthy people, and clonorchiasis, paragonimiasis were 100.0%, 91.7%, 0, 0 and 0 respectively, but the positive rates of SEA ELISA were 100.0%, 98.3%, 6.7%, 10.0% and 20.0%. The positive rates of 30 of schistosomiasis before and 4 months after treatment with P M ELISA were 100% and 70.0% respectively, with SEA ELISA were 100.0 % and 90.0%. The results suggested that the P M ELISA for detecting circulating schistosome antigen with anti rSjCTPI monoclonal antibody has higher sensitivity and specificity, as well as for evaluation of chemotherapy efficacy of schistosomiasis.