| 张永和,郁文芳,田仲文,葛继乾,陈勤生,王逸民.用酶免疫法检测细胞培养病毒的研究 I 一种简便的鉴定虫媒病毒新方法[J].Chinese journal of Epidemiology,1984,5(4):199-202 |
| 用酶免疫法检测细胞培养病毒的研究 I 一种简便的鉴定虫媒病毒新方法 |
| Using Enzyme Immunoassay For the Identification of Cell-Cultured Virus-A Simple Approach of the Identification of Arboviruses |
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| Abstract: |
| 本文报道一种十分简便的新方法，用以鉴定虫媒病毒。即用辣根过氧物酶标记金黄色葡萄球菌A蛋白作间接酶免疫测定，检测C6/36细胞培养的虫媒病毒。35株从三带喙库蚊分离的病毒，经鉴定结果20株为流行性乙型脑炎（乙脑）病毒。此与免疫荧光及蚀斑中和试验的结果完全一致。还研究了本方法的适宜条件，检测病毒滴度的敏感性以及操作的安全性等问题。结果表明本方法很值得在虫媒病毒的调查与研究中推广应用。 |
| English Abstract: |
| This paper introduces a simple and new method for the identification and testing the titration ot arbovirus isolates. Horseradish peroxidase conjugated Staphylococcus aureus protein A was used for the indirect staining of virus antigens grown in an Aedes albopictus cell line C6/36 cells.Thirty-five isolates from Culex tritaeniorhynchus mosquitoes were examined by this method and 20 of them identified as Japanese encephalitis virus (JEV). These results were confirmed by immuno-fluorescent assay and plaque neutralization test. Comparative titration of JEV, Murray Valley encephalitis and Kunjin viruses showed that this method was as sensitive as chick embryo plaque assay.Specific enzymatic reactions on these virus-infected cells began to appear before the third day and reached the end-point on the fifth day post infection of C6/36 cells, whereas the cytopathic effect appeared about 2 days later than the specific enzymatic reactions.Fixation with 10% formalin for 1/2~8 hrs did not damage the specific reaction in infected cells, nor increase the background colour for uninfected cells. |
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