CAJ | 학술논문

目的：制备基因工程表达的狂犬病毒糖蛋白优势表位抗原，并评价其在疫苗免疫后中和抗体检测中的应用价值。方法：TRIzol法从狂犬病疫苗中提取总RNA，经RT-PCR获得糖蛋白目的基因片段，构建相应的原核表达质粒，转化大肠杆菌HB101，诱导表达获得纯化重组蛋白，以重组蛋白作为包被抗原，初步建立检测糖蛋白中和抗体的ELISA方法。结果：获得狂犬病毒糖蛋白优势表位区段抗原，建立了糖蛋白中和抗体ELISA检测方法。该检测方法对59例健康献血员血浆样本检测特异性为98.31%（58/59），接种狂犬疫苗免疫个体血浆样本抗体阳性率为98.95%（94/95）。结论：基因工程表达的狂犬病毒糖蛋白优势表位抗原可用于人接种狂犬疫苗后疫苗免疫效果评价。
목적：제비기인공정표체적광견병독당단백우세표위항원，병평개기재역묘면역후중화항체검측중적응용개치。방법：TRIzol법종광견병역묘중제취총RNA，경RT-PCR획득당단백목적기인편단，구건상응적원핵표체질립，전화대장간균HB101，유도표체획득순화중조단백，이중조단백작위포피항원，초보건립검측당단백중화항체적ELISA방법。결과：획득광견병독당단백우세표위구단항원，건립료당단백중화항체ELISA검측방법。해검측방법대59례건강헌혈원혈장양본검측특이성위98.31%（58/59），접충광견역묘면역개체혈장양본항체양성솔위98.95%（94/95）。결론：기인공정표체적광견병독당단백우세표위항원가용우인접충광견역묘후역묘면역효과평개。
Objective: To clone and express a dominant epitope fragment of rabies glycoprotein and evaluate its application in neutralizing antibody detection. Methods: The dominant epitope fragment of glycoprotein was analy?ses using Biosun software and its coding gene was amplified from rabies vaccine strain by RT-PCR. After expres?sion of this dominant epitope fragment in E.coli HB101, purification was carried out in Ni-Sepharose Fast Flow columns and Sephadex G-50. Indirect ELISA system was assembled by coating the plates with the purified pro?tein. Results: SDS-PAGE results showed that dominant epitope 1~200 aa fragment was expressed efficiently in in?clusion body. ELISA results showed that system was successfully established for neutralizing antibody detection in immunized serums with positive ratio of 98.95% and a specificity of 98.31%. Conclusion: The genetic engineering dominant epitope fragment of glycoprotein can be a promising antigen in neutralizing antibody detection to make sure the successful vaccine immunization.