禽流感病毒H7亚型快速检测方法建立及应用
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摘要
通过杂交瘤技术及单克隆抗体技术获得针对禽流感病毒H7亚型的特异性单克隆抗体.抗体用于标记荧光微球,并基于双抗体夹心免疫层析法用于对禽流感病毒H7亚型的检测.针对禽流感病毒H7亚型,检出限为0.125 HAU,具有较高的灵敏度,所建立的禽流感病毒H7亚型荧光检测试纸具有良好的特异性和准确性,可用于对禽流感病毒H7亚型的现场快速检测.
The specific monoclonal antibodies against avian influenza virus H7 subtype were obtained through hybridoma technology and monoclonal antibody technology.The antibodies were used to label fluorescent microspheres and detect the avian influenza virus H7 subtype based on sandwich lateral flow immunoassay.For detecting the avian influenza virus H7 subtype,the limit of detection was 0.125 HAU with a high sensitivity.The immunoassay strips for detecting avian influenza virus H7 subtype have a good specificity and accuracy,and can be used for rapid on-site detection of the avian influenza virus H7 subtype.
The specific monoclonal antibodies against avian influenza virus H7 subtype were obtained through hybridoma technology and monoclonal antibody technology.The antibodies were used to label fluorescent microspheres and detect the avian influenza virus H7 subtype based on sandwich lateral flow immunoassay.For detecting the avian influenza virus H7 subtype,the limit of detection was 0.125 HAU with a high sensitivity.The immunoassay strips for detecting avian influenza virus H7 subtype have a good specificity and accuracy,and can be used for rapid on-site detection of the avian influenza virus H7 subtype.
引文
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[18] 吴峰,吕琦,毛茅,等.抗甲型流感病毒H1N1亚型单克隆抗体制备及应用[J].云南大学学报:自然科学版,2017(S1):106-110.
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[2] Gao R,Cao B,Hu Y,et al.Human infection with a novel avian-origin influenza A (H7N9) virus[J].New England Journal of Medicine,2013,368(20):1888.
[3] 魏茂提,赵国平,高宏伟,等.H7N9禽流感病例流行病学特征初步分析[J].武警后勤学院学报:医学版,2013(5):351-355.
[4] 孙阳,沈银忠,林萍,等.新型禽流感:H7N9[J].检验医学,2013(9):739-44.
[5] Li Q,Zhou L,Zhou M,et al.Epidemiology of human infections with avian influenza A(H7N9) virus in China[J].N Engl J Med,2014,370(6):520-532.
[6] Chen Y,Liang W,Yang S,et al.Human infections with the emerging avian influenza A H7N9 virus from wet market poultry:clinical analysis and characterisation of viral genome[J].Lancet,2013,381(9881):1 916-1 925.
[7] Uyeki T M,Cox N J.Global concerns regarding novel influenza A (H7N9) virus infections[J].New England Journal of Medicine,2013,368(20):1862.
[8] Li Q,Zhou L,Zhou M,et al.Preliminary report:Epidemiology of human infections with avian influenza A (H7N9) outbreak in China[J].New England Journal of Medcicine,2014,370(6):520-532.
[9] Xie Z,Pang YS,Liu J,et al.A multiplex RT-PCR for detection of type A influenza virus and differentiation of avian H5,H7,and H9 hemagglutinin subtypes[J].Molecular and Cellular probes,2006,20(3/4):245-249.
[10] Corman V M,Eickmann M,Landt O,et al.Specific detection by real-time reverse-transcription PCR assays of a novel avian influenza A(H7N9) strain associated with human spillover infections in China[J].Eurosurveillance,2013,18(16):10-16.
[11] Shi J Z ,Deng G H ,Liu P H ,et al.Isolation and characterization of H7N9 viruses from live poultry markets—Implication of the source of current H7N9 infection in humans[J].Chinese Science Bulletin,2013,58(16):1 857-1 863.
[12] Nie K,Zhao X,Ding X,et al.Visual detection of human infection with influenza A (H7N9) virus by subtype-specific reverse transcription loop-mediated isothermal amplification with hydroxynaphthol blue dye[J].Clinical Microbiology & Infection,2013,19(8):372-375.
[13] Wang L C,Huang D.Highly sensitive H7N9 avian influenza virus detection using reverse transcription loop-mediated isothermal amplification and oligonucleotide microarray[J].Taiwan Veterinary Journal,2015,41(4):251-255.
[14] Wong C K,Zhu H,Li OT,et al.Molecular detection of human H7N9 influenza A virus causing outbreaks in China[J].Clinical Chemistry,2013,59(7):1 062-1 07.
[15] Yu Y,Chen Z,Jian W,et al.Ultrasensitive electrochemical detection of avian influenza A (H7N9) virus DNA based on isothermal exponential amplification coupled with hybridization chain reaction of DNAzyme nanowires[J].Biosensors & Bioelectronics,2015,64:566-571.
[16] 秦智锋,吕建强,肖性龙,等.禽流感H5、H7、H9亚型多重实时荧光RT-PCR检测方法的建立[J].病毒学报,2006(2):131-136.
[17] 王秋泉,黄平,胡守旺,等.建立禽流感H7和H9亚型荧光定量PCR检测方法[J].第三军医大学学报,2010(3):246-249.
[18] 吴峰,吕琦,毛茅,等.抗甲型流感病毒H1N1亚型单克隆抗体制备及应用[J].云南大学学报:自然科学版,2017(S1):106-110.
[19] Wu F,Mao M,Cen Y,et al.Copolymerization of Eu(TTA)3Phen doped styrene and methyl methacrylate nanoparticles and use in quantitative detection of pepsinogen[J].Rsc Advances,2017,7(20):12 217-12 223.