用ELISA方法检测埃可病毒感染的特异性IgM抗体

Establishment of an indirect enzyme-linked immunosorbent assay for detecting the specific IgM antibodies in Patients with echovirus infection

目的 研究埃可病毒感染的无菌性脑膜炎的诊断方法。方法 采用埃可病毒混合抗原包被酶标板 ,用抗人γ链处理人脑脊液标本 ,通过酶标二抗及底物显色 ,建立了抗埃可病毒IgM的间接ELISA检测方法 ;并用特异性试验、对照和重复性试验证实方法的可靠性和实用性。结果 在临床诊断为无菌性脑膜炎的 78例患者脑脊液中有 14例阳性 (17 9% ) ,而细菌性脑膜炎的 36例患者脑脊液中仅有 1例阳性 (2 8% ) ,2 8例脑外伤患者脑脊液均为阴性。ELISA阳性的 5份脑脊液中和试验 4例阳性 ,而ELISA阴性的 5份脑脊液中和试验均为阴性 ;该方法与脊髓灰质炎病毒、柯萨奇A组病毒 7型和柯萨奇B组病毒 1～ 6型无交叉反应 ;ELISA阳性的 6份标本经特异性IgM破坏和阻断试验均全部转为阴性。结论 本方法快速、简便、可靠 。

Objective To provide a simple, specific and early serodiagnostic technique for the patients with aseptic meingitis caused by echovirus. Methods An indirect enzyme-linked immunosorbent assay (ELISA) has been developed to detect echovirus-IgM and the specificity and availability of the assay were also examined. Results In 78 cerebrospinal fluid (CSF) specimens which came from the children with aseptic meningitis, the positive rate was 17.9(14/78). In 64 CSF collected from non-aseptic meningitis (baterial meningitis and cerebral trauma), the positive rate was 1.56(1/64). In 5 CSF specimens which were ELISA positive, the positive rate of neutralization test (NT) was 4/5, all the specimens which were ELISA negative were NT negative. In this assay there was no cross-reaction with poliovirus, Coxsackie virus B type 1-6 and A type 7. By blocking and destructive test of specific IgM, all CSF specimens with ELISA positive became negative. Conclusion The established indirect ELISA was specific and reliable. The test was quick, simple and available, which is suitable for early and specific clinical diagnosis, and will be greatly significant to clinical treatment.