适合早期诊断的非洲猪瘟sIgA抗体量子点免疫层析检测方法建立

doi:10.11843/j.issn.0366-6964.2023.10.027

Abstract

Abstract: African swine fever (ASF), caused by the African swine fever virus (ASFV), is a devastating infectious disease in wild boars and domestic pigs. The extremely high infectivity and morbidity of ASFV have caused enormous socioeconomic losses. Given that there is no effective vaccine available for ASFV infection, early diagnosis is crucial for the prevention and control of ASF. Therefore, it is of great significance to develop a sensitive and rapid method for the on-site detection of ASFV. Firstly, the detection probe of quantum dot microspheres (QDM) coupled ASFV recombinant antigen p30 and the quality control probe of QDM coupled anti-chicken IgY were prepared based on the condensation reaction between amino and carboxyl groups. Then, the immunostrip was prepared by immobilized anti-porcine IgA-Sc antibody and chicken IgY protein on the detection line and control line, respectively. The ASFV-specific sIgA antibody in the oral fluid can be captured by the QDM-p30 fixed on the T-line. Finally, the detection sensitivity was tested by detecting the ASFV-positive oral fluid with double gradient dilution; the specificity was verified by detecting several other common swine disease viruses; and the clinical diagnostic efficacy was evaluated by testing clinical oral fluid samples from ASFV-negative and ASFV-positive animals. The proposed method shows good specificity and no cross-reaction with PRV, CSFV and PRRV; shows high sensitivity with the lowest detected dilution of 1:64 for ASFV-positive oral fluid; shows short time consuming within 20 minutes; and is easy to operate without intrusive sampling. In addition, sIgA-positive conversion could be detected as early as the 6th day after ASFV infection, suggesting the potential for early diagnosis. Here, we developed a QDM-based immunostrip for on-site early detection of ASFV sIgA antibody in oral fluid, which provides new technical support and supplement for ASFV monitoring, prevention and control.

Key words: African swine fever, sIgA antibody, QDMs-based immunostrip, point-of care testing

CLC Number:

S858.285.3

XIE Qingyun, YI Weijie, LI Jiahao, BAI Yun, XIE Xing, YUAN Ting, ZHANG Yue, FENG Yufan, ZHAO Dongming, BU Zhigao, LIU Fei, FENG Zhixin. Development of Quantum Dot Microsphere-based Immunostrip for Early Detection of Specific sIgA Antibody to African Swine Fever Virus[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4311-4319.

References 24

[1]	聂赟彬, 乔娟. 非洲猪瘟发生对我国生猪产业发展的影响[J]. 中国农业科技导报, 2019, 21(1):11-17.NIE Y B, QIAO J. Impact of African swine fever on the development of pig industry in China[J]. Journal of Agricultural Science and Technology, 2019, 21(1):11-17. (in Chinese)
[2]	COSTORD S, MUR L, LUBROTH J, et al. Epidemiology of African swine fever virus[J]. Virus Res, 2013, 173(1):191-197.
[3]	ALEJO A, MATAMOROS T, GUERRA M, et al. A proteomic atlas of the African swine fever virus Particle[J]. J Virol, 2018, 92(23):e01293-18.
[4]	SÁNCHEZ-CORDÓN P J, MONTOYA M, REIS A L, et al. African swine fever:a re-emerging viral disease threatening the global pig industry[J]. Vet J, 2018, 233:41-48.
[5]	DIXON L K, SUN H, ROBERTS H. African swine fever[J]. Antiviral Res, 2019, 165:34-41.
[6]	JAMES H E, EBERT K, MCGONIGLE R, et al. Detection of African swine fever virus by loop-mediated isothermal amplification[J]. J Virol Methods, 2010, 164(1-2):68-74.
[7]	JAUSET-RUBIO M, SVOBODOVÁ M, MAIRAL T, et al. Ultrasensitive, rapid and inexpensive detection of DNA using paper based lateral flow assay[J]. Sci Rep, 2016, 6:37732.
[8]	哈登楚日亚, 樊晓旭, 赵永刚, 等. 非洲猪瘟病毒实时荧光重组酶聚合酶扩增技术(RPA)检测方法的建立[J]. 中国畜牧兽医, 2017, 44(11):3270-3277.HADENG C R, FAN X X, ZHAO Y G, et al. Establishment of a real-time fluorescent recombinase polymerase amplification (RPA) for the detection of African swine fever virus[J]. China Animal Husbandry & Veterinary Medicine, 2017, 44(11):3270-3277. (in Chinese)
[9]	吴海涛, 成大荣, 吴萌, 等. 非洲猪瘟病毒胶体金免疫层析试纸条的研制[J]. 黑龙江畜牧兽医, 2018(17):126-128, 238.WU H T, CHENG D R, WU M, et al. Development of a colloidal gold immunochromatographic test strip for the detection of African swine fever virus[J]. Heilongjiang Animal Science and Veterinary Medicine, 2018(17):126-128, 238. (in Chinese)
[10]	上海速芯生物科技有限公司. 非洲猪瘟的快速检测——速芯科技[J]. 食品安全导刊, 2019(25):42-43.Superchip Technology. The rapid detection of African swine fever——iMole Tec[J]. China Food Safety Magazine, 2019(25):42-43. (in Chinese)
[11]	WEN X Y, XIE Q Y, LI J H, et al. Rapid and sensitive detection of African swine fever virus in pork using recombinase aided amplification combined with QDMs-based test strip[J]. Anal Bioanal Chem, 2022, 414(13):3885-3894.
[12]	GIMÉNEZ-LIROLA L G, MUR L, RIVERA B, et al. Detection of african swine fever virus antibodies in serum and oral fluid specimens using a recombinant protein 30 (P30) dual matrix indirect ELISA[J]. PLoS One, 2016, 11(9):e0161230.
[13]	REVILLA Y, PÉREZ-NÚÑEZ D, RICHT J A. African swine fever virus biology and vaccine approaches[J]. Adv Virus Res, 2018, 100:41-74.
[14]	SÁNCHEZ E, PÉREZ-NÚÑEZ D, REVILLA Y. Mechanisms of entry and endosomal pathway of african swine fever virus[J]. Vaccines (Basel), 2017, 5(4):42.
[15]	LI J H, BAI Y, LI F, et al. Rapid and ultra-sensitive detection of African swine fever virus antibody on site using QDM based-ASFV immunosensor (QAIS)[J]. Anal Chim Acta, 2022, 1189:339187.
[16]	YUAN F F, PETROVAN V, GIMENEZ-LIROLA L G, et al. Development of a blocking enzyme-linked immunosorbent assay for detection of antibodies against african swine fever virus[J]. Pathogens, 2021, 10(6):760.
[17]	WAN Y, SHI Z W, PENG G, et al. Development and application of a colloidal-gold dual immunochromatography strip for detecting African swine fever virus antibodies[J]. Appl Microbiol Biotechnol, 2022, 106(2):799-810.
[18]	ZHANG X Y, LIU X Y, WU X D, et al. A colloidal gold test strip assay for the detection of African swine fever virus based on two monoclonal antibodies against P30[J]. Arch Virol, 2021, 166(3):871-879.
[19]	SASTRE P, PEREZ T, COSTA S, et al. Development of a duplex lateral flow assay for simultaneous detection of antibodies against African and Classical swine fever viruses[J]. J Vet Diagn Invest, 2016, 28(5):543-549.
[20]	LI C F, HE X L, YANG Y, et al. Rapid and visual detection of African swine fever virus antibody by using fluorescent immunochromatography test strip[J]. Talanta, 2020, 219:121284.
[21]	王宗花, 高艳丽, 张菲菲, 等. 量子点在分析检测中的应用进展[J]. 分析科学学报, 2012, 28(1):119-125.WANG Z H, GAO Y L, ZHANG F F, et al. Applications of quantum dot in analysis and detection[J]. Journal of Analytical Science, 2012, 28(1):119-125. (in Chinese)
[22]	BAO M D, JENSEN E, CHANG Y, et al. Magnetic bead-quantum dot (MB-Qdot) clustered regularly interspaced short palindromic repeat assay for simple viral DNA detection[J]. ACS Appl Mater Interfaces, 2020, 12(39):43435-43443.
[23]	ACHADU O J, ABE F, LI T C, et al. Molybdenum trioxide quantum dot-encapsulated nanogels for virus detection by surface-enhanced raman scattering on a 2D substrate[J]. ACS Appl Mater Interfaces, 2021, 13(24):27836-27844.
[24]	BAI Y, GAN Y, HUA L Z, et al. Application of a sIgA-ELISA method for differentiation of Mycoplasma hyopneumoniae infected from vaccinated pigs[J]. Vet Microbiol, 2018, 223:86-92.

Development of Quantum Dot Microsphere-based Immunostrip for Early Detection of Specific sIgA Antibody to African Swine Fever Virus

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 24

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	BAI Yun, XIE Qingyun, OUYANG Wei, GAN Yuan, YUAN Ting, ZHAO Dongming, BU Zhigao, SHAO Guoqing, FENG Zhixin. Establishment of a Serological Method for Early Detection of African Swine Fever Virus Infection Based on Mucosal sIgA Antibody [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 300-310.
[2]	LIU Chuanxia, WANG Xiao, LI Xuewen, BAO Miaofei, LI Tingting, CHEN Xin, WENG Changjiang, ZHENG Jun. Preparation of Monoclonal Antibody of African Swine Fever Virus pE120R [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 388-394.
[3]	FENG Yongzhi, GONG Ting, WU Dongdong, GAO Qi, ZHENG Xiaoyu, ZHANG Guihong, SUN Yankuo. Analysis of Factors Affecting the Infectivity of African Swine Fever Virus on Cultured Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3406-3414.
[4]	LIU Taoxue, SU Bingqian, QI Yanli, GUO Jiangtao, LIU Zhonghu, CHU Beibei, WANG Jiang, ZENG Lei. Preparation of the Monoclonal Antibody against the African Swine Fever Virus p30 Protein and Identification of the Antigenic Epitope [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3415-3423.
[5]	DING Xiaoyan, HE Jiuxiang, ZHOU Xiaoyang, ZHOU Yuxin, LI Jintao. Preliminary Identification of Host Regulatory Genes and Virulence Genes during African Swine Fever Virus Infection [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 2964-2971.
[6]	WANG Ying, ZHU Jiahong, ZHAO Jiakai, JI Pinpin, CHEN Xu, ZHANG Lu, LIU Baoyuan, SUN Yani, ZHAO Qin. Screening and Identification of Nanobodies against NP419L Protein of African Swine Fever Virus and Its Preliminary Application of Antibody Detection [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2509-2520.
[7]	LIU Wenhao, ZHU Yance, ZHANG Dongxuan, WANG Zhihao, ZHANG Chao. Construction of PK 15 Cell Line Stably Expressing African Swine Fever Virus E165R Protein [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2662-2666.
[8]	WANG Guochao, ZHAO Yaru, ZHANG Zhonghui, ZHANG Yulong, BAI Ge, GENG Shuxian, FAN Jie, YANG Jifei, GUAN Guiquan, YIN Hong, LUO Jianxun, NIU Qingli. Bioinformatics Analysis of RNA Polymerase Subunit D205R Gene of African Swine Fever Virus and Polyclonal Antibody Preparation [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2042-2049.
[9]	ZHANG Ting, FENG Tao, YANG Jinke, HAO Yu, YANG Xing, ZHANG Dajun, SHI Xijuan, YAN Wenqian, CHEN Lingling, LIU Xiangtao, ZHENG Haixue, ZHANG Keshan. Construction and Growth Characteristics of Recombinant African Swine Fever Virus with Conditional Deletion of D1133L Gene [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(2): 706-714.
[10]	ZHANG Fangyuan, YANG Dawei, QIU Deyang, JIANG Guoqian, LI Guimei, SHAN Hu. Expression of ASFV P30 Protein and Development of ASFV Antibody Detection Method Based on x-MAP Technology [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4300-4310.
[11]	CHANG Hao, QIU Yingwu, PENG Jie, GAO Qi, SONG Zebu, CHEN Yang, LI Wei, LIN Limiao, CAO Xuezhen, ZHOU Qingfeng, ZHANG Guihong, LI Qunhui, ZHENG Zezhong. Establishment of a Dual Real-time Fluorescence Quantitative PCR Assay for Genotype I ASFV [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4428-4432.
[12]	QI Yanli, LIU Taoxue, YU Haishen, ZHANG Chao, LU Weifei, WANG Jiang, CHU Beibei, ZHANG Gaiping. Preparation of the Monoclonal Antibody against the African Swine Fever Virus p54 Protein and Identification of the Antigenic Epitope [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 281-292.
[13]	WANG Yang, CUI Shuai, XIN Ting, WANG Xixi, YU Hainan, CHEN Shiyu, JIANG Yajun, GAO Xintao, PANG Zhongbao, JIANG Yitong, GUO Xiaoyu, JIA Hong, ZHU Hongfei. ASFV MGF360-14L Interacts with MAVS and Inhibit the Expression of Type Ⅰ Interferon [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(9): 3272-3278.
[14]	ZHAO Xuyang, JIN Jiaxin, LU Wenlong, ZHANG Shuai, HUANG Li, ZHANG Gaiping, SUN Aijun, ZHUANG Guoqing. Advances in the Molecular Mechanism of Immune Escape of African Swine Fever Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(7): 2074-2082.
[15]	CHEN Xiaojun, MA Jun, CHEN Xiongnan, LIANG Yifan, GAO Qi, HUANG Zhao, XU Runda, ZHENG Jiachen, ZHENG Zezhong, ZHANG Guihong, WANG Heng, GONG Lang. Identification of the B Cell Epitopes Recognized by a Monoclonal Antibody against the p30 Protein of African Swine Fever Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(7): 2239-2251.