现场快速检测大田软海绵酸的传感器与仪器的研究
|
摘要
研究并建立针对大田软海绵酸OA(okadaic acid)快速、有效的酶联免疫吸附试验ELISA(Enzyme-Linked ImmunosorbentAssay)定量检测方法,制备了定量检测试剂盒-生物酶传感器,并研发配套的现场快速检测仪器-仿生电子眼(Bionic e-Eye),实现对OA的快速检测。对标准样品的参数进行测定,该OA试剂盒的检测限为0.19μg/L,检测范围为0.2μg/L~5μg/L;样品回收率为75%~125%,精密度在±15%以内。将Bionic e-Eye的检测结果和酶标仪及高效液相色谱HPLC(high performance liquid chromatog-raphy)所测数据的结果进行了比较,结果表明,Bionic e-Eye的性能与酶标仪的检测结果一致。此外,该仪器可以直接得出检测结果,不需要人工计算处理,适用于现场的快速检测,将是一种新的现场快速定量检测OA的技术手段。
Based on Enzyme-linked immunosorbent assay(ELISA),test kit,namely,enzyme sensor and the corre-sponding instrument was prepared,Bionic e-Eye for rapid,effective and quantitative detection of okadaic acid(OA)on-site. The detection limit of the kit is 0.19 μg/L and the range is among 0.2 μg/L and 5 μg/L. The recovery rate isamong 75% and 125% and the accuracy is within±15%. The performance of Bionic e-Eye is consistent with micro-plate reader and high performance liquid chromatography(HPLC). In addition,without artificial calculation,theinstrument can show the results directly on the screen,which is very suitable for rapid detection,and will be a newtechnology for rapid and quantitative detection of OA on site.
Based on Enzyme-linked immunosorbent assay(ELISA),test kit,namely,enzyme sensor and the corre-sponding instrument was prepared,Bionic e-Eye for rapid,effective and quantitative detection of okadaic acid(OA)on-site. The detection limit of the kit is 0.19 μg/L and the range is among 0.2 μg/L and 5 μg/L. The recovery rate isamong 75% and 125% and the accuracy is within±15%. The performance of Bionic e-Eye is consistent with micro-plate reader and high performance liquid chromatography(HPLC). In addition,without artificial calculation,theinstrument can show the results directly on the screen,which is very suitable for rapid detection,and will be a newtechnology for rapid and quantitative detection of OA on site.
引文
[1]Dominguez R B,Hayat A,Sassolas A,et al.Automated FlowThrough Amperometric Immunosensor for Highly Sensitive andon-Line Detection of Okadaic Acid in Mussel Sample[J].Talanta,2012,99:232-237.
[2]Hayat A,Barthelmebs L,Marty J L.Enzyme-Linked Limmunosen-sor Based on Super Paramagnetic Nanobeads for Easy and RapidDetection of Okadaic Acid[J].Analytica Chimica Acta,2011,690(2):248-252.
[3]Ehara H,Makino M,Kodama K,et al.Crystal Structure of OkadaicAcid Binding Protein 2.1:A Sponge Protein Implicated in CytotoxinAccumulation[J].Chem Bio Chem,2015,16(10):1435-1439.
[4]Perreault F,Matias M S,Oukarroum A,et al.Okadaic acid InhibitsCell Growth and Photosynthetic Electron Transport in the AlgaDunaliella Tertiolecta[J].Science of the Total Environment,2012,414:198-204.
[5]Stengel D B,Connan S.Natural Products From Marine Algae:Methods and Protocols[M].New York:Humana Press,2015:277-297.
[6]Morton S L,Donald R T.Determination of Okadaic Acid Contentof Dinoflagellate Cells:A Comparison of the HPLC-FluorescentMethod and Two Monoclonal Antibody ELISA Test Kits[J].Toxi-con,1996,34(8):947-954.
[7]柳其芳.ELISA法测定贝毒素污染的研究[J].中国热带医学,2012,12(11):1398-1399.
[8]Sassolas A,Hayat A,Catanante G,et al.Detection of the MarineToxin Okadaic Acid:Assessing Seafood Safety[J].Talanta,2013,105:306-316.
[9]王天星,黎洪波,苏凯麒,等.基于细胞电阻抗传感器的细胞多生理参数分析系统设计[J].传感技术学报,2014,27(12):1589-1595.
[10]苏凯麒,邹玲,王琴,等.基于细胞阻抗传感器的腹泻性毒素检测系统设计与实现[J].传感技术学报,2014,27(3):283-288.
[11]Hu N,Wang T,Cao J,et al.Comparison Between ECIS and LAPSfor Establishing a Cardiomyocyte-Based Biosensor[J].Sensorsand Actuators B:Chemical,2013,185(8):238-244.
[12]Su K,Zou Q,Zhou J,et al.High-Sensitive and High-Efficient Bio-chemical Analysis Method Using a Bionic Electronic Eye in Com-bination with a Smartphone-Based Colorimetric Reader System[J].Sensors and Actuators B:Chemical,2015,216:134-140.
[13]邹瞿超,方佳如,苏凯麒,等.基于移动终端的麻痹性贝类毒素现场快速检测系统设计[J].传感技术学报,2015,28(9):1269-1274.
[2]Hayat A,Barthelmebs L,Marty J L.Enzyme-Linked Limmunosen-sor Based on Super Paramagnetic Nanobeads for Easy and RapidDetection of Okadaic Acid[J].Analytica Chimica Acta,2011,690(2):248-252.
[3]Ehara H,Makino M,Kodama K,et al.Crystal Structure of OkadaicAcid Binding Protein 2.1:A Sponge Protein Implicated in CytotoxinAccumulation[J].Chem Bio Chem,2015,16(10):1435-1439.
[4]Perreault F,Matias M S,Oukarroum A,et al.Okadaic acid InhibitsCell Growth and Photosynthetic Electron Transport in the AlgaDunaliella Tertiolecta[J].Science of the Total Environment,2012,414:198-204.
[5]Stengel D B,Connan S.Natural Products From Marine Algae:Methods and Protocols[M].New York:Humana Press,2015:277-297.
[6]Morton S L,Donald R T.Determination of Okadaic Acid Contentof Dinoflagellate Cells:A Comparison of the HPLC-FluorescentMethod and Two Monoclonal Antibody ELISA Test Kits[J].Toxi-con,1996,34(8):947-954.
[7]柳其芳.ELISA法测定贝毒素污染的研究[J].中国热带医学,2012,12(11):1398-1399.
[8]Sassolas A,Hayat A,Catanante G,et al.Detection of the MarineToxin Okadaic Acid:Assessing Seafood Safety[J].Talanta,2013,105:306-316.
[9]王天星,黎洪波,苏凯麒,等.基于细胞电阻抗传感器的细胞多生理参数分析系统设计[J].传感技术学报,2014,27(12):1589-1595.
[10]苏凯麒,邹玲,王琴,等.基于细胞阻抗传感器的腹泻性毒素检测系统设计与实现[J].传感技术学报,2014,27(3):283-288.
[11]Hu N,Wang T,Cao J,et al.Comparison Between ECIS and LAPSfor Establishing a Cardiomyocyte-Based Biosensor[J].Sensorsand Actuators B:Chemical,2013,185(8):238-244.
[12]Su K,Zou Q,Zhou J,et al.High-Sensitive and High-Efficient Bio-chemical Analysis Method Using a Bionic Electronic Eye in Com-bination with a Smartphone-Based Colorimetric Reader System[J].Sensors and Actuators B:Chemical,2015,216:134-140.
[13]邹瞿超,方佳如,苏凯麒,等.基于移动终端的麻痹性贝类毒素现场快速检测系统设计[J].传感技术学报,2015,28(9):1269-1274.