抗前列腺素D合成酶自身抗体酶联免疫吸附试验检测方法的建立及其在不育症诊断中的初步应用

Establishment of enzyme-linked immunosorbent assay for anti-prostaglandin D synthase autoantibody and its preliminary application for infertility diagnosis

目的 :证实抗 lipocalin型前列腺素 D合成酶 ( L -PGDS)自身抗体的存在 ,建立检测该抗体的酶联免疫吸附试验 ( EL ISA)方法 ,并探讨其在不育症诊断中的初步应用。方法 :用纯化的重组 L-PGDS作为抗原 ,建立检测抗 L-PGDS自身抗体的 EL ISA法 ,并用特异性 L -PGDS吸附试验检测 EL ISA法的特异性 ,同时检测 EL ISA法的重复性 ,并对2 0 4例不育症病人的精浆和血清中抗 L-PGDS自身抗体进行检测。结果 :特异性吸附试验和重复性试验证实 ,EL ISA法稳定可靠 ,1 1 4例不育男性中精浆抗 L-PGDS自身抗体阳性者 49例 ( 3 4.0 % ) ;无精子症、少精子症病人的阳性率 ( 48.5%和 43 .2 % )明显高于正常精子数的不育男性 ( 2 3 .0 % ) ( P<0 .0 1和 P<0 .0 5)。 60例不育病人血清抗 L-PGDS自身抗体阳性 1 8例 ( 3 0 .0 % )。结论 :不育症病人精浆和血清中存在抗 L-PGDS自身抗体 ,建立的 EL ISA法稳定可靠 。

Objective: To confirm existence of anti-lipocalin-type prostaglandin D synthase (L-PGDS) autoantibody in seminal plasma and serum, an enzyme-linked immunosorbent assay (ELISA) was established. Its application in infertility diagnosis was also evaluated. Methods: ELISA was established to detect anti-L-PGDS autoantibody by using purified recombinant L-PGDS as antigen and its specificity and repetition were evaluated. The anti-L-PGDS autoantibody in seminal plasma and serum from 204 cases of infertile patients was detected by this method. Results: ELISA was confirmed to be stable and reliance by specific sorption and repetition test. There were 49 cases (34.0%) of seminal plasma positive anti-L-PGDS autoantibody in 114 cases of infertile men. The positive rate of seminal plasma anti-L-PGDS autoantibody in aspermia patients (48.5%) or oligospermia patients (43.2%) was significantly higher than that in infertile patients with normal sperm density (23.0%) (P<0.01 and P<0.05 respectively). There were 18 cases (30.0%) of serum positive anti-L-PGDS autoantibody in 60 cases of infertile patients. Conclusion: There is anti-L-PGDS autoantibody in seminal plasma or serum from infertile patients. ELISA used to detect anti-L-PGDS autoantibody is stable and reliance and suitable for clinical application.