中华实验和临床病毒学杂志
中華實驗和臨床病毒學雜誌
중화실험화림상병독학잡지
CHINESE JOURNAL OF EXPERIMENTAL AND CLINICAL VIROLOGY
2009年
6期
427-429
,共3页
滕灵方%林峰%郑美云%郑昌华%吴锋%曾爱平%黄恩佩%莫一涵%郑敏巧%李旭阳%侯建毅
滕靈方%林峰%鄭美雲%鄭昌華%吳鋒%曾愛平%黃恩珮%莫一涵%鄭敏巧%李旭暘%侯建毅
등령방%림봉%정미운%정창화%오봉%증애평%황은패%막일함%정민교%리욱양%후건의
细小病毒科%基因表达调控%免疫印迹法
細小病毒科%基因錶達調控%免疫印跡法
세소병독과%기인표체조공%면역인적법
Parvovifidae%Gene expression profding%lmmunoblotting
目的 表达人博卡病毒(HBoV)VP_2蛋白,用于临床标本HBoV的筛查.方法 将VP_2基因重组于杆状病毒基因组,重组的杆状病毒感染昆虫细胞sf9,用HA抗体进行Western Blot鉴定重组融合VP_2蛋白表达.通过电镜观察重组蛋白颗粒.以VP_2蛋白作为抗原对176例临床样本进行免疫印迹筛查.结果 在昆虫细胞中VP_2蛋白的表达量约占细胞总蛋白的60%以上,VP_2蛋白相对分子质量为60×10~3.电镜观察博卡病毒VP_2蛋白形成病毒样颗粒(VLP),其大小在20mm左右;采用免疫印迹技术从临床标本中筛查出4例患者HBoV阳性,阳性率达到2.28%(4/176).结论 本研究成功的在昆虫细胞中表达了博卡病毒VP_2蛋白;表达VP_2蛋白具有一定抗原性,为开展人博卡病血清学研究方法建立提供基础.
目的 錶達人博卡病毒(HBoV)VP_2蛋白,用于臨床標本HBoV的篩查.方法 將VP_2基因重組于桿狀病毒基因組,重組的桿狀病毒感染昆蟲細胞sf9,用HA抗體進行Western Blot鑒定重組融閤VP_2蛋白錶達.通過電鏡觀察重組蛋白顆粒.以VP_2蛋白作為抗原對176例臨床樣本進行免疫印跡篩查.結果 在昆蟲細胞中VP_2蛋白的錶達量約佔細胞總蛋白的60%以上,VP_2蛋白相對分子質量為60×10~3.電鏡觀察博卡病毒VP_2蛋白形成病毒樣顆粒(VLP),其大小在20mm左右;採用免疫印跡技術從臨床標本中篩查齣4例患者HBoV暘性,暘性率達到2.28%(4/176).結論 本研究成功的在昆蟲細胞中錶達瞭博卡病毒VP_2蛋白;錶達VP_2蛋白具有一定抗原性,為開展人博卡病血清學研究方法建立提供基礎.
목적 표체인박잡병독(HBoV)VP_2단백,용우림상표본HBoV적사사.방법 장VP_2기인중조우간상병독기인조,중조적간상병독감염곤충세포sf9,용HA항체진행Western Blot감정중조융합VP_2단백표체.통과전경관찰중조단백과립.이VP_2단백작위항원대176례림상양본진행면역인적사사.결과 재곤충세포중VP_2단백적표체량약점세포총단백적60%이상,VP_2단백상대분자질량위60×10~3.전경관찰박잡병독VP_2단백형성병독양과립(VLP),기대소재20mm좌우;채용면역인적기술종림상표본중사사출4례환자HBoV양성,양성솔체도2.28%(4/176).결론 본연구성공적재곤충세포중표체료박잡병독VP_2단백;표체VP_2단백구유일정항원성,위개전인박잡병혈청학연구방법건립제공기출.
Objective To clone and express VP_2 gene from HBoV,and the expressed VP_2 protein was as the antigen in order to detect serum from children in Wenling area with lower respiratory tract infections.Methods The VP_2 gene was reeombined with the genome of Baculovirus,which infected the insect cell.The fusion protein with HA tag was applied to confirm the specificity of expressed protein.Furthermore,the recombinant protein was observed using electron microscopy.The 176 serum from children in Wenling area with lower respiratory tract infections was screened using Western blot.Results The expressed VP_2 protein was more than 60% in total proteins from insect cell,and MWt about 60 ×10~3.The virus-like particle(VLP)Was observed using electron microscopy,and size about 20nm.The 176 serum from children in wenling area with lower respiratory tract infections was screened using Western blot.The HBoV positive rate Was 2.28%(4/176).Conclusion The VP_2 protein from human bocavims was expressed in insect cell successfully.Through HA tag the VP_2 protein wag specific,and then the assay using SDS-PAGE with Western blot could detect and screen the antibody in serum from children with lower respiratory tract infections rapidly and accurately.
