基于花状硫化铜和纳米抗体的免疫层析试纸条法检测黄曲霉毒素B_(1)  

Detection of aflatoxin B_(1)by immunochromatographic test strip based on CuS nanoflowers and nanobody

作　　者：赵一鸣 宋子尧 何保山[1] 任文洁 ZHAO Yiming;SONG Ziyao;HE Baoshan;REN Wenjie(College of Food Science and Engineering,National Engineering Laboratory/Key Laboratory of Henan Province,Henan University of Technology,Zhengzhou 450001,China)

机构地区：[1]河南工业大学粮油食品学院,河南工业大学国家工程实验室/省重点实验室,河南郑州450001

出　　处：《河南工业大学学报(自然科学版)》2024年第6期82-89,共8页Journal of Henan University of Technology:Natural Science Edition

基　　金：河南工业大学粮油食品学科国家工程实验室/省重点实验室开发课题基金资助项目(NC2021013);河南省高等学校重点科研项目(22A550004)。

摘　　要：黄曲霉毒素B_(1)(AFB_(1))是有毒代谢产物,不仅污染农作物,还容易通过食物链进入人体,影响人类健康。为了及时、快速、方便地检测出农作物中的AFB_(1),需要一种低成本、用户友好、高效的检测方法。构建以硫化铜纳米花(CuS Nanoflowers,CuS NFs)为信号标签,纳米抗体G8-DIG为识别探针的免疫层析试纸条法用于检测黄曲霉毒素B_(1)。对CuS NFs的质量浓度、G8-DIG的添加量、T线质量浓度、探针量进行优化。结果表明:该方法的检出限(LOD)为2.4 ng/mL,检测范围为6.87~96.21 ng/mL,加标回收率在86%~110%之间,变异系数(CV)≤10.9%;对玉米质控样进行检测,回收率为111.8%。选用的CuS NFs和G8-DIG作为信号探针,能够在检测AFB_(1)的同时降低试纸条的成本。Aflatoxin B_(1)(AFB_(1))is a toxic metabolite typically produced by Aspergillus flavus and parasitic fungi,not only contaminates crops but also readily enters the human body through the food chain,affecting human health.Given the widespread distribution,severe contamination,significant hazards to humans and animals,as well as the difficulty in removal of AFB_(1),there is an urgent need for a convenient and sensitive detection method.In contrast to instrumental analysis methods,immunochromatographic test strip(ICTS)offers advantages such as short detection times,high efficiency,and no requirement for specialized personnel.In view of the wide applications of ICTS and the excessive variety and quantity of crops tested,it is necessary to reduce the cost of ICTS.Nanobody G8 was fused with an anti-digoxin antibody to construct the expression vector pET25b-G8-DIG,which was then transformed into E.coli BL21 for prokaryotic expression.Following nickel column purification,the synthesized G8-DIG fusion protein was confirmed to have a molecular weight of approximately 30 kDa through SDS-PAGE gel electrophoresis.The multifunctional fusion protein G8-DIG was able to bind to both AFB_(1)antigen and digoxin(DIG)antigen.Copper sulfide nanoflowers(CuS Nanoflowers,CuS NFs)were synthesized by the reaction of copper nitrate trihydrate and thiourea,and the nanomaterial was confirmed to possess a floral structure with dimensions around 300 nm through TEM and SEM analysis.Optimization of the ICTS including the optimal mass concentration of CuS NFs solution,the amount of G8-DIG added,the mass concentration of the T-line,and the probe quantity revealed that the detection limit(LOD)of this method was 2.4 ng/mL,with a detection range of 6.87-96.21 ng/mL.The spiked recovery rate ranged from 86%to 110%,and the coefficient of variation(CV)was less than 10.9%.When tested on corn quality control samples,the recovery rate was 111.8%.Furthermore,the specificity of the method was assessed by detecting OTA,T-2,DON,and ZEN as interfering samples on t

关 键 词：硫化铜纳米花 纳米抗体 免疫层析试纸条 黄曲霉毒素B_(1) 

分 类 号：TS207.3[轻工技术与工程—食品科学]