国际医学寄生虫病杂志
國際醫學寄生蟲病雜誌
국제의학기생충병잡지
INTERNATIONAL JOURNAL OF MEDICAL PARASITIC DISEASES
2011年
3期
129-134
,共6页
陈军虎%王越%坪井敬文%韩银泽
陳軍虎%王越%坪井敬文%韓銀澤
진군호%왕월%평정경문%한은택
间日疟原虫%体液免疫%蛋白芯片
間日瘧原蟲%體液免疫%蛋白芯片
간일학원충%체액면역%단백심편
Plasmodium vivax%Humoral immune%Protein chip
目的 建立检测间日疟感染的Ni-NTA蛋白芯片技术.方法 采用无细胞蛋白合成体系表达问日疟原虫重组蛋白,建立原位纯化重组蛋白和检测间日疟原虫感染患者血清中抗体反应的Ni-NTA蛋白芯片技术.并对3个裂殖子表面蛋白(merozoite surface proteins,MSPs)MSPl-42、MSP8和MSP10的免疫应答进行分析.结果 应用Ni-NTA蛋白芯片技术检测问日疟原虫感染患者血清抗体,鉴定出具有免疫原性的15个间日疟原虫蛋白,主要包括10个MSPs、2个Cys6蛋白以及其他3个未知蛋白,结果与以往报道的抗体芯片类似.MSPl-42、MSP8和MSP10依次识别出100.0%(20/20)、90.0%(18/20)和70.0%(14/20)的间日疟原虫感染患者血清,特异性均为100%(10/10),且曲线下面积(area under the curve.AUC)达到0.87～1.00.结论 成功建立了检测间日疟原虫感染的Ni-NTA蛋白芯片技术.该方法有助于从间日疟原虫基因组中快速筛选鉴定具有免疫原性的蛋白以及应用于功能蛋白质组学研究.
目的 建立檢測間日瘧感染的Ni-NTA蛋白芯片技術.方法 採用無細胞蛋白閤成體繫錶達問日瘧原蟲重組蛋白,建立原位純化重組蛋白和檢測間日瘧原蟲感染患者血清中抗體反應的Ni-NTA蛋白芯片技術.併對3箇裂殖子錶麵蛋白(merozoite surface proteins,MSPs)MSPl-42、MSP8和MSP10的免疫應答進行分析.結果 應用Ni-NTA蛋白芯片技術檢測問日瘧原蟲感染患者血清抗體,鑒定齣具有免疫原性的15箇間日瘧原蟲蛋白,主要包括10箇MSPs、2箇Cys6蛋白以及其他3箇未知蛋白,結果與以往報道的抗體芯片類似.MSPl-42、MSP8和MSP10依次識彆齣100.0%(20/20)、90.0%(18/20)和70.0%(14/20)的間日瘧原蟲感染患者血清,特異性均為100%(10/10),且麯線下麵積(area under the curve.AUC)達到0.87～1.00.結論 成功建立瞭檢測間日瘧原蟲感染的Ni-NTA蛋白芯片技術.該方法有助于從間日瘧原蟲基因組中快速篩選鑒定具有免疫原性的蛋白以及應用于功能蛋白質組學研究.
목적 건립검측간일학감염적Ni-NTA단백심편기술.방법 채용무세포단백합성체계표체문일학원충중조단백,건립원위순화중조단백화검측간일학원충감염환자혈청중항체반응적Ni-NTA단백심편기술.병대3개렬식자표면단백(merozoite surface proteins,MSPs)MSPl-42、MSP8화MSP10적면역응답진행분석.결과 응용Ni-NTA단백심편기술검측문일학원충감염환자혈청항체,감정출구유면역원성적15개간일학원충단백,주요포괄10개MSPs、2개Cys6단백이급기타3개미지단백,결과여이왕보도적항체심편유사.MSPl-42、MSP8화MSP10의차식별출100.0%(20/20)、90.0%(18/20)화70.0%(14/20)적간일학원충감염환자혈청,특이성균위100%(10/10),차곡선하면적(area under the curve.AUC)체도0.87～1.00.결론 성공건립료검측간일학원충감염적Ni-NTA단백심편기술.해방법유조우종간일학원충기인조중쾌속사선감정구유면역원성적단백이급응용우공능단백질조학연구.
Objective To develop Ni-NTA surface based chips to detect the humoral immune response to Plasmodium vivax infection.Methotis A set of recombinant P.vivax proteins was expressed by wheat germ cell-free system and IgG immune responses to these recombinant P. vivax proteins including 3 merozoite surface proteins(MSPl-42,MSP8 and MSPIO)were measured with sera from P.vwax-infected patients and healthv individuals using Ni-NTA chips.Results Ni-NTA surface based chips were successfully applied to detect immune responses to P.vivaz infection.Fifteen high immunoreactive recombinant P.vivax proteins were identified using Ni-NTA chips,including 10 MSPs,2 Cys6 lipid raft-associated proteins and 3 hypothetical proteins.MSPI-42.MSP8 and MSPIO recognized by IgG immune response to 100.0%(20/20),90.0%(18/20),and 70.0%(14/20)serum samples from P.vivax infected patients,respectively.Moreover,no crossreactivity to MSPs was found in serum samples from healthy individuals with area under the curve(AUC)values of 0.87-1.00.Conclusions In this report,Ni-NTA chips have been applied to investigate blood stage-specific immunogenic proteins from vivax malaria.The method may aid in determining the immunogenicity of candidate antigens and the functional identification of the large number of unknown and hypothetical proteins in P.vivax genome.
