微生物学免疫学进展
微生物學免疫學進展
미생물학면역학진전
PROGRESS IN MICROBIOLOGY AND IMMUNOLOGY
2014年
5期
28-32
,共5页
许笑笑%钟建琴%张文捷%邵铭%刘书珍%苏波%李长贵%姚伟
許笑笑%鐘建琴%張文捷%邵銘%劉書珍%囌波%李長貴%姚偉
허소소%종건금%장문첩%소명%류서진%소파%리장귀%요위
四价流感病毒裂解疫苗(QIV)%B型血凝素含量%单向免疫扩散法%抗原%参考品
四價流感病毒裂解疫苗(QIV)%B型血凝素含量%單嚮免疫擴散法%抗原%參攷品
사개류감병독렬해역묘(QIV)%B형혈응소함량%단향면역확산법%항원%삼고품
Quadrivalent influenza vaccine (split virion) inactivated ( QIV)%B type HA content%Single radial diffusion ( SRD)%Antigen%Standard
目的:建立能准确测定四价流感病毒裂解疫苗中B型血凝素含量(单向免疫扩散法,SRD)的检测方法。方法采用双价抗原参考品SRD法对四价流感病毒裂解疫苗中两种B型血凝素含量进行测定。将B1与B2抗原参考品按质量浓度1∶1混合制备为双价抗原参考品,双价抗原和待检样品与10%裂解剂按9∶1比例裂解30 min,分别加入到含有抗血清参考品的1.5%琼脂糖凝胶板上,每孔10μL,置20~25℃放置18~24 h,经干燥、染色、脱色,测量结果。验证双价抗原SRD法的重复性和准确性。结果双价抗原SRD法测定的血凝素含量平均值比单价抗原SRD法更接近理论配制值,故双价抗原SRD法比单价抗原SRD法能更能准确检测QIV中两种B型血凝素含量,经验证双价抗原SRD法的重复性及准确性良好。结论双价抗原SRD法提高了四价流感病毒裂解疫苗B型血凝素含量检测的准确性,为精确测定四价流感病毒裂解疫苗中B型血凝素含量提供了有效的方法和数据支持。
目的:建立能準確測定四價流感病毒裂解疫苗中B型血凝素含量(單嚮免疫擴散法,SRD)的檢測方法。方法採用雙價抗原參攷品SRD法對四價流感病毒裂解疫苗中兩種B型血凝素含量進行測定。將B1與B2抗原參攷品按質量濃度1∶1混閤製備為雙價抗原參攷品,雙價抗原和待檢樣品與10%裂解劑按9∶1比例裂解30 min,分彆加入到含有抗血清參攷品的1.5%瓊脂糖凝膠闆上,每孔10μL,置20~25℃放置18~24 h,經榦燥、染色、脫色,測量結果。驗證雙價抗原SRD法的重複性和準確性。結果雙價抗原SRD法測定的血凝素含量平均值比單價抗原SRD法更接近理論配製值,故雙價抗原SRD法比單價抗原SRD法能更能準確檢測QIV中兩種B型血凝素含量,經驗證雙價抗原SRD法的重複性及準確性良好。結論雙價抗原SRD法提高瞭四價流感病毒裂解疫苗B型血凝素含量檢測的準確性,為精確測定四價流感病毒裂解疫苗中B型血凝素含量提供瞭有效的方法和數據支持。
목적:건립능준학측정사개류감병독렬해역묘중B형혈응소함량(단향면역확산법,SRD)적검측방법。방법채용쌍개항원삼고품SRD법대사개류감병독렬해역묘중량충B형혈응소함량진행측정。장B1여B2항원삼고품안질량농도1∶1혼합제비위쌍개항원삼고품,쌍개항원화대검양품여10%렬해제안9∶1비례렬해30 min,분별가입도함유항혈청삼고품적1.5%경지당응효판상,매공10μL,치20~25℃방치18~24 h,경간조、염색、탈색,측량결과。험증쌍개항원SRD법적중복성화준학성。결과쌍개항원SRD법측정적혈응소함량평균치비단개항원SRD법경접근이론배제치,고쌍개항원SRD법비단개항원SRD법능경능준학검측QIV중량충B형혈응소함량,경험증쌍개항원SRD법적중복성급준학성량호。결론쌍개항원SRD법제고료사개류감병독렬해역묘B형혈응소함량검측적준학성,위정학측정사개류감병독렬해역묘중B형혈응소함량제공료유효적방법화수거지지。
Objective To establish the HA content determination method ( SRD) of type B for quadrivalent influenza vac-cine(QIV).Methods The determination of hemagglutinin (HA) content for two types of B strains in QIV was performed by using of a bivalent reference antigen .Prepare bivalent reference antigen with mix B 1 and B2 reference antigen according to 1 ∶1(C/C).Split bivalent reference antigen and samples with 10%split solution in 9 ∶1 ratio for 30 min, load 10μL/hole samples to 1.5%agarose gel that with a enough specific reference antiserum .Then put that gel at 20-25 ℃for 18~24 hours, then drying, staining, destaining and measure he result .Validate the repeatability and accuracy of the bivalent reference SRD method .Results The mean is more close to theoretical value by using bivalent reference than using mono-valent reference to determine B type of QIV .So it is more accurate by using bivalent reference than using monovalent refen-rence to test HA of type B .The accuracy and repeatability of this method is satisfied demonstrated by validating .Conclu-sion The developed method could improve accuracy in determination of type B strains in QIV .It approve effective method and supported data to determinate HA content of type B for QIV .