Radioimmunofluorescent antibody technique for detection of reovirus antigen in cell culture
Radioimmunofluorescent antibody technique for detection of reovirus antigen in cell culture
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DOI：
10.1128/iai.14.3.811-815.1976
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发表时间：
1976-09
影响因子：
3.1
通讯作者：
J. R. McCammon
中科院分区：
文献类型：
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作者：
J. R. McCammon
抗呼肠孤病毒I型抗体必须部分纯化并与异硫氰酸荧光素（FITC）偶联。然后将FITC标记的抗体与125I缀合。正常血清的丙种球蛋白组分同样用FITC标记，然后用131 I标记。然后，将FITC +125 I标记的免疫试剂与FITC +131 I对照试剂在相同蛋白质基础上混合。该混合物用于染色丙酮固定的呼肠孤病毒感染的盖玻片。染色后，通过荧光显微镜检查盖玻片。感染的盖玻片表现出呼肠孤病毒的特征性免疫荧光，而未感染的盖玻片为阴性。目视检查后，将盖玻片置于试管中，并在双通道伽马分析仪中计数。通过将定量同位素数据与来自单一制剂的免疫荧光的定性信息进行比较，可以将抗原产生位点与定量产生值相关联。
Antibody against reovirus type I must partially purified and conjugated with fluorescein isothiocyanate (FITC). The FITC-labeled antibody was then conjugated with 125I. A gamma globulin fraction of normal sera was similarly labeled with FITC followed by a 131I label. The FITC + 125I-labeled immune reagent was then mixed on an equal protein basis with the FITC + 131I control reagent. This mixture was used to stain acetone-fixed reovirus-infected cover slips. After staining, the cover slips were examined by fluorescence microscopy. Infected cover slips demonstrated characteristic reovirus immunofluorescence, whereas uninfected cover slips were negative. After visual examination, the cover slips were placed in tubes and counted in a two-channel gamma analyzer. By comparing the quantitative isotope data with the qualtitative information from immunofluorescence on a single preparation, it was possible to correlate antigen production sites with quantitative production values.