两种方法检测EHF患者早期血清中病毒结构蛋白抗体的研究

DETECTIONOFANTIBODIESAGAINSTSTRUCTURAL PROTEINS OFEHFVINTHESERAOFTHEEARLY PATIENTSWITHTWOMETHODS

ＥＨＦ病毒抗原经已知抗核蛋白（ＮＰ）的ＭｃＡｂＡ３５及抗膜蛋白的ＭｃＡｂＡ３Ｇ１亲和层析进行分离纯化，再经高压液相色谱（ＨＰＬＣ）进一步纯化，获得了高纯度的ＥＨＦ病毒结构蛋白：ＮＰ、ＭＰ。书ＮＰ、ＭＰ分别滴于ＰＶＤＦ膜，与早期患者血清进行免疫酶斑点试验，检测患者血清中相应结构蛋白的抗体，并结合临床资料进行临床转归分析；同时与ＨＩ、ＲＰＨＩ检测结果相比较。２８例ＥＨＦ患者血清，斑点试验检测仅ＭＰ抗体阳性４例，ＨＩ检出３例；ＲＰＨＩ检测仅ＮＰ抗体阳性１５例，有５例斑点试验同时检出ＭＰ抗体，说明斑点试验对ＭＰ抗体的检测敏感性高（抗体滴度亦高）。联系ＭＰ、ＮＰ抗体的出现与病情转归的关系发现，ＭＰ抗体先出现的患者病情普遍转重，ＮＰ抗体先出现的患者，病情转轻或趋于稳定，ＭＰ、ＮＰ抗体同时出现的患者，其转归介于上述二者之间，且ＨＩ、ＲＰＨＩ和斑点试验检测结果其差别无统计意义。提示斑点试验用于ＥＨＦ患者血清中结构蛋白抗体的检测更敏感，有可能作为患者病情转归的预测指标，值得进一步推广应用。

EHFV antigens were isolated and purified by affinity chromatography andhigh pressure liquid chromatog raphy with momoclonal antibodies angainst nucleocapsidprotein(NP)and membrane protein(MP).After purified ,NP, MP were dropped in PVDFmembrane to carry out immune enzyme dot assay in order to detect corresponding antibodiesinthe sera of EHF patients. Simutaneously the results of dot assay compared with that ofHI,RPHI. Of 28 sera of EHF early cases; 3 were positive by HI in 4 sera only detected MPantibody by dot assay ;5 were at the same time detected out MP antibody by dot assay in 1 5sera only detecting NP antibody by RPHI ,which proved that dot assay detecting MPantibody was more sensitive than Hl·The most patients first detected NP antibody becamebetter or stabilization , but the patients first detected M P antibody became generally worse. Both HI ,RPHI and dot assay had the same results(p>0.05).This study showed thatimmune enzyme dot assay could be used to detect NP , M P antibodies in the sera of EHFpatients and forecast patient cover ,more specific, sensitive ,reliable than HI ,RPHI.