埃博拉病毒抗体间接ELISA检测方法的建立及应用

doi:10.11843/j.issn.0366-6964.2016.03.027

Abstract

Abstract:

An indirect ELISA which can detect EBOV antibody was established using the purified the envelope glycoprotein as coating antigen.We use the HRP labeled antibody as the secondary antibody，the positive and negative horse sera as controls.After the reaction conditions were optimized，we analyzed the specificity，sensitivity of the indirect ELISA using serum of horse which immunized with EBOV virus like particles.For further evaluation of application of the ELISA，the results were compared to the neutralization assay based on pseudo-type EBOV.The indirect ELISA could detect the antibody against EBOV specifically and showed no cross-reaction with the positive sera of West Nile Fever et al.The indirect ELISA also showed high coincidence with the neutralization assay based on pseudo-type EBOV in evaluation of the immune response after immunization.The coefficient of variation of inter-batch and intra-batch duplicative tests was less than 8％.The indirect ELISA was successfully developed.It provides an effective and convenient method to kinetic detection of antibody against EBOV after immunization.

CLC Number:

S852.659.5

CAO Zeng-guo，WANG Hua-lei，GAI Wei-wei，ZHENG Xue-xing，JIN Hong-li，LI Ling，WANG Li-na，FENG Na，ZHAO Yong-kun，WANG Tie-cheng，GAO Yu-wei，BI Yu-hai，YANG Song-tao，XIA Xian-zhu. Development of an Indirect ELISA for Detecting Ebola Virus Antibody and Its Application[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(3): 615-619.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.xmsyxb.com/EN/10.11843/j.issn.0366-6964.2016.03.027

https://www.xmsyxb.com/EN/Y2016/V47/I3/615

References

［1］林祥梅，韩雪清，王景林.外来动物疫病［M］.北京：科学出版社，2014：296-324.
LIN X M，HAN X Q，WANG J L.Exotic animal disease［M］.Beijing：Science Press，2014：296-324.(in Chinese)
［2］殷震，刘景华.动物病毒学［M］.北京：科学出版社，1997：773-776.
YIN Z，LIU J H.Animal virology［M］.Beijing：Science Press，1997：773-776.(in Chinese)
［3］LI H，YING T，YU F，et al.Development of therapeutics for treatment of Ebola virus infection［J］.Microbes Infect，2015，17(2)：109-117.
［4］扈荣良.现代动物病毒学［M］.北京：中国农业出版社，2014：1029-1039.
HU R L.Modern animal virology［M］.Beijing：China Agriculture Press，2014：1029-1039.(in Chinese)
［5］程颖，刘军，李昱，等.埃博拉病毒病：病原学、致病机制、治疗与疫苗研究进展［J］.科学通报，2014，59(30)：2889-2899.
CHENG Y，LIU J，LI Y，et al.Ebola virus disease：virology，pathogenesis，therapy，and vaccines［J］.Chinese Science Bulletin，2014，59(30)：2889-2899.(in Chinese)
［6］金宏丽，王化磊，郑学星，等.埃博拉病毒病疫苗研究进展［J］.传染病信息，2015，28(2)：70-74.
JIN H L，WANG H L，ZHENG X X，et al.Advances in the development of Ebloa vaccines［J］.Infectious Disease Information，2015，28(2)：70-74.(in Chinese)
［7］郑学星，杨松涛，王化磊，等.埃博拉疫苗研究新进展［J］.中国病原生物学杂志，2013，8(7)：652-655.
ZHENG X X，YANG S T，WANG H L，et al.Recent advances in Ebola virus vaccine development［J］.Journal of Pathogen Biology，2013，8(7)：652-655.(in Chinese)
［8］SAIJO M，NIIKURA M，MORIKAWA S，et al.Enzyme-linked immunosorbent assays for detection of antibodies to Ebola and Marburg viruses using recombinant nucleoproteins［J］. J Clin Microbiol，2001，39(1)：1-7.
［9］PREHAUD C，HELLEBRAND E，COUDRIER D，et al.Recombinant Ebola virus nucleoprotein and glycoprotein(Gabon 94 strain) provide new tools for the detection of human infections［J］.J Gen Virol，1998，79(Pt 11):2565-2572.
［10］乔晋娟，罗俊，危宏平.埃博拉病毒感染的实验室诊断方法研究进展［J］.科技导报，2015，33(1)：100-106.
QIAO J J，LUO J，WEI H P.Laboratory diagnosis of Ebola virus disease：A review［J］.Science & Technology Review，2015，33(1)：100-106.(in Chinese)
［11］MARTIN P，LAUPLAND K B，FROST E H，et al.Laboratory diagnosis of Ebola virus disease［J］.Intensive Care Med，2015，41(5)：895-898.
［12］王颖芳，段广才.埃博拉病毒及其致病性［J］.新乡医学院学报，2014，31(11)：877-882.
WANG Y F，DUAN G C.Ebola virus and its pathogenicity［J］.Journal of Xinxiang Medical University，2014，31(11)：877-882.(in Chinese)
［13］MANICASSAMY B，WANG J，JIANG H，et al.Comprehensive analysis of ebola virus GP1 in viral entry［J］.J Virol，2005，79(8)：4793-4805.
［14］WILLET M，KURUP D，PAPANERI A，et al.Preclinical development of inactivated rabies virus-based polyvalent vaccine against rabies and filoviruses［J］.J Infect Dis，2015，212(Suppl 2):414-424.
［15］NIIKURA M，IKEGAMI T，SAIJO M，et al.Detection of Ebola viral antigen by enzyme-linked immunosorbent assay using a novel monoclonal antibody to nucleoprotein［J］.J Clin Microbiol，2001，39(9)：3267-3271.

[1]	MENG Lingzhai, CHEN Chunli, YU Mengmeng, WANG Zhanxin, WANG Suyan, LIU Peng, HE Tana, GUO Ru, CHEN Yuntong, LIU Changjun, QI Xiaole, WU Zhiqiang, GAO Yulong. Pathogenicity of Avian Metapneumovirus Subtype B on Yellow Feather Broilers and Evaluation on Immune Effect of Inactivated Vaccine [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 5154-5161.
[2]	JIANG Shengqiang, HU Jing, CHEN Hongying. Expression Analysis of CircRNAs in A549 Cells Infected with H1N1 Influenza A Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4724-4734.
[3]	ZHANG Ao, TAN Bin, LIU Kexin, LIU Jiali, ZHANG Shuqin. Genomic Characterization Analysis of a H1N1 Subtype Swine Influenza Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4866-4871.
[4]	HE Chenxiang, GAO Shandian, TIAN Zhancheng, DU Junzheng, WANG Jinming, GUAN Guiquan, YIN Hong. Development of a Differential Diagnostic ELISA Based on G_NS Protein to Distinguish BEFV Infected and Vaccinated Cattle [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4320-4326.
[5]	CHANG Yiming, TANG Cheng, YUE Hua. Molecular Epidemiological Investigation of Bovine Respiratory Syncytial Virus in Yaks [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2030-2041.
[6]	FANG Yuan, HOU Qiaodi, XIANG Chaohui, ZHAO Hongyi, QI Xuefeng. Regulatory Effects of IFITM3 on Proliferation of Peste des Petits Ruminants Virus (PPRV) in Goat Endometrial Epithelial Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2200-2207.
[7]	LI Shun, LI Guizhen, YUAN Yaoxian, LI Kangjian, MA Chunquan, LI Shoujun, HUANG Liangzong, MA Jun. Effect of the Length of PA-X Gene on Replication and Pathogenicity of H3N2 Canine Influenza Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 272-280.
[8]	CUI Mingxian, WANG Xingbo, HUANG Yanming, BIAN Xiyi, FENG Mengke, YAN Yan, DONG Weiren, ZHOU Jiyong. Genetic Characterization and Evolution of Three Strains of H3N2 Avian Influenza Viruses [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(11): 4116-4122.
[9]	YU Mengmeng, BAO Yuanling, WANG Suyan, XIN Ziqi, LIU Peng, FENG Xiaoyan, MENG Lingzhai, GUO Ru, ZHANG Yanping, LIU Changjun, QI Xiaole, LI Junping, WANG Xiaomei, GAO Yulong. Isolation, Identification and Pathogenicity of Subtype B Avian Metapneumovirus [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(10): 3540-3549.
[10]	ZHOU Yong, LI Zhixin, LU Hongwei, SUN Yan, LI Tian, DU Fanshu, PU Juan. Surveillance and Outbreak Analysis of H5 and H7N9 Subtypes of Highly Pathogenic Avian Influenza in China [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(9): 3093-3106.
[11]	PAN Chunrong, DENG Ruixue, HU Linjie, ZHU Xueliang, SUN Yuefeng, WANG Guirong, MENG Xuelian. Effect of Histone Deacetylase Inhibitors on Peste Des Petits Ruminants Virus Replication [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(7): 2307-2316.
[12]	CHEN Zixuan, ZHANG Nan, HU Qun, QUAN Keji, QIN Tao, CHEN Sujuan, PENG Daxin, LIU Xiufan. Mutation Analysis of Antigen Sites 145 and 153 on Hemagglutinin of H9N2 Avian Influenza Virus in China [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(4): 1165-1172.
[13]	DUAN Zhiqiang, XIE Lingling, CHEN Jiaqi, TANG Hong, WANG Yanbi, ZHAO Caiqin, ZHAO Jiafu. Research Progress on the Interactions of Newcastle Disease Virus M Protein with Host Proteins [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(1): 32-42.
[14]	YAN Yayao, GU Min, LIU Xiufan. Advance in the Influence of Amino Acid Variation in HA Protein on the Biological Properties of H7N9 Subtype Influenza Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(8): 2093-2106.
[15]	JIANG Mei, CHEN Wu, ZHAI Junqiong, BU Wandi, XIE Yilun, LIU Canbin, SHAN Fen, LUO Manlin. Cloning and Sequence Analysis of H and F Genes of Canine Distemper Virus Strain from Red Panda [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(6): 1670-1676.

Development of an Indirect ELISA for Detecting Ebola Virus Antibody and Its Application

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments