山东农业科学
山東農業科學
산동농업과학
SHANGDONG AGRICULTURAL SCIENCES
2014年
4期
102-105
,共4页
房保海%贾俊涛%梁旭锋%姜英辉%李正义%房倩%金莹
房保海%賈俊濤%樑旭鋒%薑英輝%李正義%房倩%金瑩
방보해%가준도%량욱봉%강영휘%리정의%방천%금형
赭曲霉毒素A(OTA)%量子点%双抗夹心荧光免疫检测(sFLISA)
赭麯黴毒素A(OTA)%量子點%雙抗夾心熒光免疫檢測(sFLISA)
자곡매독소A(OTA)%양자점%쌍항협심형광면역검측(sFLISA)
Ochratoxin A (OTA)%Quantum dots (QDs)%Sandwich fluorescence -linked immunosor-bent assay (sFLISA)
建立了基于量子点标记技术的赭曲霉毒素A(Ochratoxin A,OTA)双抗夹心荧光免疫检测(sand-wich fluorescence -linked immunosorbent assay ,sFLISA)体系,包含多克隆包被抗体、生物素标记的多克隆检测抗体、量子点标记的链霉亲和素等,获得了sFLISA的最佳操作参数:包被抗体浓度2.5μg/mL,检测抗体稀释500倍,量子点标记链霉亲和素稀释100倍。该方法在OTA浓度3.125~125μg/L之间时,相对荧光强度和OTA浓度呈线性关系,回归方程为y=0.0206x+0.2018,R2=0.9924;加标回收率在90.1%~110.0%之间,变异系数均小于10%,能较好地进行赭曲霉毒素A的定量检测。
建立瞭基于量子點標記技術的赭麯黴毒素A(Ochratoxin A,OTA)雙抗夾心熒光免疫檢測(sand-wich fluorescence -linked immunosorbent assay ,sFLISA)體繫,包含多剋隆包被抗體、生物素標記的多剋隆檢測抗體、量子點標記的鏈黴親和素等,穫得瞭sFLISA的最佳操作參數:包被抗體濃度2.5μg/mL,檢測抗體稀釋500倍,量子點標記鏈黴親和素稀釋100倍。該方法在OTA濃度3.125~125μg/L之間時,相對熒光彊度和OTA濃度呈線性關繫,迴歸方程為y=0.0206x+0.2018,R2=0.9924;加標迴收率在90.1%~110.0%之間,變異繫數均小于10%,能較好地進行赭麯黴毒素A的定量檢測。
건립료기우양자점표기기술적자곡매독소A(Ochratoxin A,OTA)쌍항협심형광면역검측(sand-wich fluorescence -linked immunosorbent assay ,sFLISA)체계,포함다극륭포피항체、생물소표기적다극륭검측항체、양자점표기적련매친화소등,획득료sFLISA적최가조작삼수:포피항체농도2.5μg/mL,검측항체희석500배,양자점표기련매친화소희석100배。해방법재OTA농도3.125~125μg/L지간시,상대형광강도화OTA농도정선성관계,회귀방정위y=0.0206x+0.2018,R2=0.9924;가표회수솔재90.1%~110.0%지간,변이계수균소우10%,능교호지진행자곡매독소A적정량검측。
The sandwich fluorescence -linked immunosorbent assay ( sFLISA) system was established to detect ochratoxin A (OTA), which included coated antibody , detecting antibody (labeled by biotin), strepta-vidin ( labeled by QDs ) and other elements .The optimal operating parameters were obtained:coated concen-tration was 2.5 μg/mL;dilution ratio of detecting antibody was 1∶500;dilution ratio of streptavidin (labeled by QDs) is 1∶100.There was a linear relationship between RFU and OTA concentration in the range of 3.125~125 μg/L, and the linear equation was y =0.0206x +0.2018 (R2 =0.9924).The average recovery was between 90 .1%and 110 .0%, and the coefficient of variation was below 10%.This system could detect OTA correctly.