[Objectives]This study was conducted to develop an enzyme-linked immunoassay kit that can detect the residual amount of pentachloronitrobenzene in Penaeus vannamei.[Methods]This study was conducted to develop an enzym...[Objectives]This study was conducted to develop an enzyme-linked immunoassay kit that can detect the residual amount of pentachloronitrobenzene in Penaeus vannamei.[Methods]This study was conducted to develop an enzyme-linked immunoassay kit that can detect the residual amount of pentachloronitrobenzene in P.vannamei.[Results]The standard curve range of the kit was 0-8.1μg/L;the detection limit for P.vannamei was 0.912μg/kg;the recovery was 80.6%-103.5%;and the relative standard deviation range within batches was 5.3%-10.1%,and the relative standard deviation range between batches was 6.7%-8.1%.The specificity of the pentachloronitrobenzene monoclonal antibody was relatively good,and the cross-reaction rates with pentachlorophenol,hexachlorobenzene,tetrachlorophthalide,and chlorothalonil were low,all of which did not exceed 30%.The ELISA kit could be stored at 4℃for 12 months,showing good stability.[Conclusions]The detection kit has low cost,short time and small deviation,and is an ideal preliminary screening method.展开更多
目的建立肠道病毒71型(enterovirus type 71,EV-A71)灭活疫苗(Vero细胞)抗原含量检测方法并对其进行方法学验证及初步应用。方法以抗EV-A71兔多克隆抗体为包被抗体,辣根过氧化物酶(horseradish peroxidase,HRP)标记的抗EV-A71鼠单克隆...目的建立肠道病毒71型(enterovirus type 71,EV-A71)灭活疫苗(Vero细胞)抗原含量检测方法并对其进行方法学验证及初步应用。方法以抗EV-A71兔多克隆抗体为包被抗体,辣根过氧化物酶(horseradish peroxidase,HRP)标记的抗EV-A71鼠单克隆抗体为检测抗体,建立双抗体夹心ELISA并验证其线性范围、专属性、准确度、精密度及耐用性等。通过检测EV-A71疫苗原液及研发生产工艺各中间制品的EV-A71抗原含量评价该方法的适用性。结果包被抗体与酶标抗体最佳稀释度分别为1∶5000和1∶10000。抗原为5~80 U/mL时,线性良好;与同为肠道病毒的其他抗原不存在交叉反应,专属性良好;对不同浓度抗原进行6次测定,其回收率在90%~100%,准确度良好;不同实验员对不同浓度抗原样品检测3次,CV均<11%,精密度良好;针对不同影响因素设计耐用性试验,回收率均在80%~120%,耐用性良好。该方法检测EV-A71疫苗原液和制备过程中的中间制品均具有良好的线性和平行性,表明该方法具有良好的适用性。结论建立了EV-A71疫苗(Vero细胞)抗原含量检测方法并进行了初步应用,为EV-A71疫苗和HFMD多价疫苗的研发及质量控制提供了可靠的检测方法。展开更多
基金Key R&D Program of Hebei Province:Special Project on Key Common Technologies for High-quality Agricultural Development(20327505D).
文摘[Objectives]This study was conducted to develop an enzyme-linked immunoassay kit that can detect the residual amount of pentachloronitrobenzene in Penaeus vannamei.[Methods]This study was conducted to develop an enzyme-linked immunoassay kit that can detect the residual amount of pentachloronitrobenzene in P.vannamei.[Results]The standard curve range of the kit was 0-8.1μg/L;the detection limit for P.vannamei was 0.912μg/kg;the recovery was 80.6%-103.5%;and the relative standard deviation range within batches was 5.3%-10.1%,and the relative standard deviation range between batches was 6.7%-8.1%.The specificity of the pentachloronitrobenzene monoclonal antibody was relatively good,and the cross-reaction rates with pentachlorophenol,hexachlorobenzene,tetrachlorophthalide,and chlorothalonil were low,all of which did not exceed 30%.The ELISA kit could be stored at 4℃for 12 months,showing good stability.[Conclusions]The detection kit has low cost,short time and small deviation,and is an ideal preliminary screening method.
文摘目的建立肠道病毒71型(enterovirus type 71,EV-A71)灭活疫苗(Vero细胞)抗原含量检测方法并对其进行方法学验证及初步应用。方法以抗EV-A71兔多克隆抗体为包被抗体,辣根过氧化物酶(horseradish peroxidase,HRP)标记的抗EV-A71鼠单克隆抗体为检测抗体,建立双抗体夹心ELISA并验证其线性范围、专属性、准确度、精密度及耐用性等。通过检测EV-A71疫苗原液及研发生产工艺各中间制品的EV-A71抗原含量评价该方法的适用性。结果包被抗体与酶标抗体最佳稀释度分别为1∶5000和1∶10000。抗原为5~80 U/mL时,线性良好;与同为肠道病毒的其他抗原不存在交叉反应,专属性良好;对不同浓度抗原进行6次测定,其回收率在90%~100%,准确度良好;不同实验员对不同浓度抗原样品检测3次,CV均<11%,精密度良好;针对不同影响因素设计耐用性试验,回收率均在80%~120%,耐用性良好。该方法检测EV-A71疫苗原液和制备过程中的中间制品均具有良好的线性和平行性,表明该方法具有良好的适用性。结论建立了EV-A71疫苗(Vero细胞)抗原含量检测方法并进行了初步应用,为EV-A71疫苗和HFMD多价疫苗的研发及质量控制提供了可靠的检测方法。
