摘要
建立了一种基于生物条形码结合杂交链式反应扩增的高灵敏、特异性检测蓖麻毒素(Ricin toxin,RT)的方法。使用RT多克隆抗体(p Ab)及条形码DNA (Barcode DNA)制备功能性金纳米探针(GNPs),通过与RT单克隆抗体(m Ab)共同识别作用,形成"m Ab-RT-pAb/Au NPs/Barcode DNA"三明治夹心结构,进一步利用条形码DNA作为引发链触发生物素标记的发卡探针进行杂交链式扩增,形成一条具有重复单元的长双链DNA。当使用辣根过氧化物酶标记链霉亲和素与长链DNA上的生物素结合后,通过鲁米诺与过氧化氢的化学发光底物发光进行测定。结果表明,本方法检测RT的线性范围为0.61 ng/m L～5.00μg/m L,检出限为0.52 ng/m L(S/N=3),本方法具有较宽的检测范围及较低的检出限,测定饮用水及脱脂牛奶样品中RT的加标回收率在83.4%～112.4%之间,相对标准偏差在2.2%～5.4%之间,表明本方法实用性良好,在食品和饮用水安全控制及防范生物恐怖袭击方面具有良好的应用前景。
A novel highly sensitive and specific method for detection of ricin toxin(RT)based on bio-barcode combined with hybridization chain reaction(HCR)amplification was developed.Functional gold nanoparticles(GNPs)were coated with anti-ricin polyclonal antibodies(pAb)and barcode DNA.The"mAb-RT-pAb/AuNPs/barcode DNA"sandwich structure was formed by co-identification with the anti-ricin monoclonal antibody(mAb),and HCR was further carried out by using barcode DNA as an initiation chain probe for triggering the chain-triggered biotin label,which resulted in a long double-stranded DNA with repeating units.Immediately,streptavidin labeled horseradish peroxidase(SA-HRP)was combined with biotin on long strand DNA.Finally,the chemiluminescence values were detected through adding substrates of luminol and hydrogen peroxide.The results showed that the linear range of RT was 0.61 ng/mL-5000 ng/mL,and the limit of detection(LOD)was 0.52 ng/mL(S/N=3).This method showed a wide detection range and a low detection limit.The recoveries of spiked samples in milk and water samples ranged from 83.4%to 112.4%,and the relative standard deviations(RSDs)ranged from 2.2%to 5.4%.This method exhibited great application prospects in food drinking water safety control and prevention of bioterrorism attacks.
作者
苑帅
霍冰洋
张曼
王瑜
白家磊
彭媛
宁保安
陈萍
高志贤
YUAN Shuai;HUO Bing-Yang;ZHANG Man;WANG Yu;BAI Jia-Lei;PENG Yuan;NING Bao-An;CHEN Ping;GAO Zhi-Xian(Food Science and Engineering College,Jilin Agricultural University,Changchun 130118,China;Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Institute of Environmental and Operational Medicine, Academy of Military Medical Science,Academy of Military Science,Tianjin 300050,China)
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2019年第1期147-154,共8页
Chinese Journal of Analytical Chemistry
基金
国家重点研发计划项目(No.2017YFC1200903)
天津市科技支撑重点项目(No.18YFZCNC01260)资助~~
