Study of enteric non-A, non-B hepatitis virus

我们尝试通过免疫印迹、免疫电子显微镜和基因工程技术对肠道非甲、非乙型肝炎病毒及其蛋白进行鉴定。对肠道性非甲、非乙型肝炎患者的粪便进行过滤，滤液进行超速离心。取上清液和微丸，用10%嘧达胺凝胶进行凝胶电泳法。考马斯亮蓝蛋白染色可见100kD、、26kodalton条带。然而，通过使用恢复期患者的血清进行蛋白质印迹分析，没有发现特定的条带。光镜下，肠道型非甲、非乙型肝炎的组织学表现与其他急性肝炎基本一致。经电子显微镜观察，变性肝细胞胞浆内可见病毒样颗粒。免疫电子显微镜检查不能将其鉴定为肠道非甲非乙型肝炎病毒颗粒。从患者粪便中提取核糖核酸(RNA)，反转录成c DNA，与lambda GT 11载体连接。用患者恢复期血清对lambda GT11文库进行筛选，未获得阳性克隆。虽然我们用聚合酶链式反应的方法对这些RNA进行了检测，但也没有得到阳性结果，我们无法获得阳性结果的原因可能是材料不好，不足以提取病毒RNA，或者是因为抗体产生得太弱，无法检测到病毒蛋白。目前正在进行进一步检查，以从其他患者的粪便材料和恢复期血清中鉴定病毒核酸和病毒蛋白。

We have tried to identify the enteric non-A, non-B hepatitis virus and their proteins by western blotting, immune-electron microscopy, and genetic engineering techniques.1 . Fecal materials from patients with enteric non-A, non-B hepatitis were filtered and the filtrants were ultracentrifuged. Then both supernatents and pellets were electrophoresed through 10% acridamide gel. By protein staining with Coomassie blue, bands were observed at size of 100, 64, and 26 kilodalton. However, no specific bands were identified by western blotting using patients' serum at their convalescent stage.2. By light microscopic examination, histological findings of enteric non-A, non-B hepatitis were almost identical with those of other acute hepatitis. By electron microscopic examination, virus-like particles were observed in the cyteplasm of degenerated hepatocytes. However, we could not identify them as enteric non-A, non-B hepatitis virus particles by immune-electron microscopic examination.3. Ribonucleic acids (RNA) were extracted from patients fecal materials, then reverse transcribed to cDNA, and ligated with lambda gt 11 vector. The lambda gt 11 library were screened with patients convalescent phase serum, but no positive clone was obtained. Although we examined these RNA by polymerase chain reaction methods, no positive findings were obtained as well.The reason why we could not obtain the positive finding may be because either the materials were not good; enough to get viral RNA or because antibody production is too weak to detect viral proteins. Further examination is now underway to identify the virus nucleic acids and viral proteins with fecal materials and convalescent phase sera from other patients.