基于SPR生物传感器的H5N1流感病毒快速检测方法
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摘要
为利用表面等离子共振(Surface Plasmon Resonance,SPR)生物传感器建立一种快速检测H5N1流感病毒的新方法。该研究利用SPR生物传感器以及H5N1单克隆抗体,对H5N1流感病毒样品进行检测分析。结果表明:该方法特性如下:1)可实现针对H5N1流感病毒的直接、快速和实时检测,整个过程无需任何标记;2)检测限为104.44 TCID50/mL,低于ELISA方法的103.24 TCID50/mL;3)在抗体消耗量方面,SPR技术要优于ELISA技术近100倍。
An efficient alternative method to detect the presence of H5 N1 virus based on surface plasmon resonance(SPR)and monoclonal antibody against Influenza A(H5 N1)virus was proposed in this study.Samples of H5 N1 influenza virus were tested and analyzed.The results showed:1)The good performance of SPR biosensor was based on the direct detection,real time record,and without any additional labeling.2)The detection limit of the assay was found to be 104.44 TCID50/mL,which was lower than the limit of ELISA of 103.24 TCID50/mL.3)The consumption of antibody in SPR test was 100-fold lower than that in ELISA test.Thus,in the future,with the development of high quality virus antigen and antibody production,the SPR biosensor system would become the most commonly applied and efficient approach for influenza A viruses diagnosis.
An efficient alternative method to detect the presence of H5 N1 virus based on surface plasmon resonance(SPR)and monoclonal antibody against Influenza A(H5 N1)virus was proposed in this study.Samples of H5 N1 influenza virus were tested and analyzed.The results showed:1)The good performance of SPR biosensor was based on the direct detection,real time record,and without any additional labeling.2)The detection limit of the assay was found to be 104.44 TCID50/mL,which was lower than the limit of ELISA of 103.24 TCID50/mL.3)The consumption of antibody in SPR test was 100-fold lower than that in ELISA test.Thus,in the future,with the development of high quality virus antigen and antibody production,the SPR biosensor system would become the most commonly applied and efficient approach for influenza A viruses diagnosis.
引文
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[3] Dhumpa R,Handberg K J,Jorgensen P H,Yi S,Wolff A,Bang D D.Rapid detection of avian influenza virus in chicken fecalsamplesbyimmunomagneticcapturereverse transcriptase-polymerase chain reaction assay[J].Diagnostic Microbiology and Infectious Disease,2011,69(3):258-265
[4] Kolosova A Y,Shim W B,Yang Z Y,Eremin S A,Chung D H.Direct competitive ELISA based on a monoclonal antibody for detection of aflatoxin B1 Stabilization of ELISA kit components and application to grain samples[J].Analytical and Bioanalytical Chemistry,2006,384(1):286-294
[5] Chen Y W,Luo W X,Song H J,Yin B Y,Tang J X,Chen Y X,Ng M H,Yeo A E T,Zhang J,Xia N S.Mimotope ELISA for detection of broad spectrum antibody against avian H5N1influenza virus[J].PLoS One,(2011-06-09):e24144DOI:10.1371/journal.pone.0024144
[6] Myszka D G,Rich R L.Implementing surface plasmon resonance biosensors in drug discovery[J].Pharmaceutical Science and Technology Today,2000,3(9):310-317
[7] Wong C L,Chua M,Mittman H,Choo L X,Lim H Q,Olivo M.A Phase-intensity surface plasmon resonance biosensor for avian influenza A(H5N1)detection[J].Sensors,2017,17(10),2363DOI:10.3390/s17102363
[8] Pennacchio A,Varriale A,Esposito M G,Scala A,Marzullo V M,Staiano M,D′auria S.A rapid and sensitive assay for the detection of benzylpenicillin(PenG)in Milk[J].PLoS One,(2015-07-13)DOI:10.1371/journal.pone.0132396
[9] Luo C,Luo H B,Zheng S X,Gui C S,Yue L D,Yu C Y,Sun T,He P L,Chen J,Shen J H,Luo X M,Li Y X,Liu H,Bai D L,Shen J K,Yang Y M,Li F Q,Zuo J P,Hilgenfeld R,Pei G,Chen K X,Shen X,Jiang H L.Nucleocapsid protein of SARS coronavirus tightly binds to human cyclophilin A[J].Biochemical and Biophysical Research Communications,2004,321(3):557-565
[10] Hearty S,Conroy P J,Ayyar B V,Byrne B,O′kennedy R.Surface plasmon resonance for vaccine design and efficacy studies:Recent applications and future trends[J].Expert Review of Vaccines,2010,9(6):645-664
[11] VaisocherovH,Faca V M,Taylor A D,Hanash S Y,Jiang S.Comparative study of SPR and ELISA methods based on analysis of CD166/ALCAM levels in cancer and control human sera[J].Biosensors and Bioelectronics,2009,24(7):2143-2148
[12] Saksono B,Dewi B E,Nainggolan L,Suda Y S.A highly sensitive diagnostic system for detecting dengue viruses using the interaction between a sulfated sugar chain and a virion[J].PLoS One,2015,10(5):e0123981DOI:10.1371/journal.pone.0123981
[13] Hu J D,Wang T T,Wang S,Chen M W,Wang M P,Mu L Y,Chen H Y,Hu X R,Liang H,Zhu J H,Jiang M.Development of a surface plasmon resonance biosensing approach for the rapid detection of porcine circovirus type2in sample solutions[J].PLoS One,(2014-10-29)DOI:10.1371/journal.pone.0111292
[14] Ashley J,D′aurelio R,Piekarska M,Temblay J,Pleasants M,Trinh L,Rodgers T,Tothill I.Development of a betaLactoglobulin sensor based on SPR for milk allergens detection[J].Biosensors,2018,8(2),32 DOI:10.3390/bios8020032
[15] Li Y B,Wang R H.Aptasensors for Detection of Avian Influenza Virus H5N1[M].In:Li Y B,Wang R H.eds Biosensors and Biodetection.New York,NY:Springer New York,2017
[16] Nguyen V T,Seo H B,Kim B C,Kim S K,Song C S,Gu M B.Highly sensitive sandwich-type SPR based detection of whole H5Nx viruses using apair of aptamers[J].Biosensors and Bioelectronics,2016,86:293-300
[17] Kang K N,Lee Y S.RNA Aptamers:A Review of Recent Trends and Applications[M].In:Kang K N,Lee Y S.eds.Advances in Biochemical Engineering/Biotechnology.Berlin,Heidelberg:Springer Berlin Heidelberg,2012