Development and evaluation of polyclonal antibodies based antigen capture ELISA for detection of porcine rotavirus

被引:4
|
作者
Memon, Atta Muhammad [1 ]
Chen, Fangzhou [1 ]
Khan, Sher Bahadar [1 ]
Guo, Xiaozhen [1 ]
Khan, Rajwali [2 ]
Khan, Farhan Anwar [1 ]
Zhu, Yinxing [1 ]
He, Qigai [1 ]
机构
[1] Huazhong Agr Univ, Coll Vet Med, State Key Lab Agr Microbiol, Wuhan, Peoples R China
[2] Univ Agr Peshawar, Dept Livestock Management Breeding & Genet, Peshawar, Pakistan
关键词
Porcine rotavirus; VP6; polyclonal antibodies; AC-ELISA; POTENTIALLY ZOONOTIC VIRUSES; GROUP-A; DIARRHEA; PIGS; ANIMALS; FECES;
D O I
10.1080/10495398.2022.2052304
中图分类号
S8 [畜牧、 动物医学、狩猎、蚕、蜂];
学科分类号
0905 ;
摘要
Rotaviruses are rising as zoonotic viruses worldwide, causing the lethal dehydrating diarrhea in children, piglets, and other livestock of economic importance. A simple, swift, cost-effective, highly specific, and sensitive antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) was developed for detection of porcine rotavirus-A (PoRVA) by employing rabbit (capture antibody) and murine polyclonal antibodies (detector antibody) produced against VP6 of PoRVA (RVA/Pig-tc/CHN/TM-a/2009/G9P23). Reactivity of the both polyclonal antibodies was confirmed by using an indirect ELISA, western-blot analysis and indirect fluorescence assay against rVP6 protein and PoRVA. The detection limit of AC-ELISA was found 50 ng/ml of PoRVA protein. The relative sensitivity and specificity of this in-house AC-ELISA were evaluated for detection of PoRVA from 295 porcine diarrhea samples, and results were compared with that of RT-PCR and TaqMan RT-qPCR. The relative sensitivity and specificity of AC-ELISA compared with those of TaqMan RT-qPCR were found as 94.4 and 99.2%, respectively, with the strong agreement (kappa -0.58) between these two techniques. Furthermore, AC-ELISA could not detect any cross-reactivity with porcine epidemic diarrhea virus, transmissible gastro-enteritis virus, pseudo rabies virus and porcine circovirus-2. This in-house AC-ELISA efficiently detected PoRVA from clinical samples, which suggests that this technique can be used for large-scale surveillance and timely detection of rotavirus infection in the porcine farms. NOVELTY STATEMENT In this study, we used a Chinese porcine rotavirus-A (PoRVA) strain containing the I5, a dominant VP6-genotype in pigs, for production of VP6 (most conserved) protein based polyclonal antibodies (pAb) in rabbits (as capture Ab) and mouse (as detector Ab) for development of simple, cost effective, highly specific and sensitive AC-ELISA for detection of PoRVA. Furthermore, there is no any previous published report on application of rabbit and mouse pAb against VP6 for developing an AC-ELISA against PoRVA.
引用
收藏
页码:1807 / 1814
页数:8
相关论文
共 50 条
  • [41] A RAPID ANTIGEN CAPTURE ELISA FOR THE DETECTION OF ORTHOPOX VIRUSES
    JOHANN, S
    CZERNY, CP
    JOURNAL OF VETERINARY MEDICINE SERIES B-ZENTRALBLATT FUR VETERINARMEDIZIN REIHE B-INFECTIOUS DISEASES AND VETERINARY PUBLIC HEALTH, 1993, 40 (08): : 569 - 581
  • [42] Development of a Novel Double Antibody Sandwich ELISA for Quantitative Detection of Porcine Deltacoronavirus Antigen
    Wang, Wei
    Li, Jizong
    Fan, Baochao
    Zhang, Xuehan
    Guo, Rongli
    Zhao, Yongxiang
    Zhou, Junming
    Zhou, Jinzhu
    Sun, Dongbo
    Li, Bin
    VIRUSES-BASEL, 2021, 13 (12):
  • [43] Production of polyclonal antibodies against ochratoxin A and its detection in chilies by ELISA
    Thirumala-Devi, K
    Mayo, MA
    Reddy, G
    Reddy, SV
    Delfosse, P
    Reddy, DVR
    JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 2000, 48 (10) : 5079 - 5082
  • [44] Development and application of an indirect ELISA for detection of antibodies against emerging atypical porcine pestivirus
    Hao Song
    Xiaowei Gao
    Jing Li
    Xinying Dong
    Yanhui Fu
    Lina Shao
    Jiaoer Zhang
    Hua-Ji Qiu
    Yuzi Luo
    Virology Journal, 21
  • [45] Development and application of an indirect ELISA for detection of antibodies against emerging atypical porcine pestivirus
    Song, Hao
    Gao, Xiaowei
    Li, Jing
    Dong, Xinying
    Fu, Yanhui
    Shao, Lina
    Zhang, Jiaoer
    Qiu, Hua-Ji
    Luo, Yuzi
    VIROLOGY JOURNAL, 2024, 21 (01)
  • [46] Development and application of a recombinant M protein-based indirect ELISA for the detection of porcine deltacoronavirus IgG antibodies
    Luo, Shang-Xing
    Fan, Jing-Hui
    Opriessnig, Tanja
    Di, Jing-Mei
    Liu, Bao-Jing
    Zuo, Yu-Zhu
    JOURNAL OF VIROLOGICAL METHODS, 2017, 249 : 76 - 78
  • [47] Development and application of an indirect ELISA for the detection of antibodies to porcine epidemic diarrhea virus based on a recombinant spike protein
    Lin, Huixing
    Zhou, Hong
    Gao, Lu
    Li, Bin
    He, Kongwang
    Fan, Hongjie
    BMC VETERINARY RESEARCH, 2018, 14
  • [48] Development and application of an indirect ELISA for the detection of antibodies to porcine epidemic diarrhea virus based on a recombinant spike protein
    Huixing Lin
    Hong Zhou
    Lu Gao
    Bin Li
    Kongwang He
    Hongjie Fan
    BMC Veterinary Research, 14
  • [49] AN ELISA FOR SULFONAMIDE DETECTION USING AFFINITY-PURIFIED POLYCLONAL ANTIBODIES
    ASSIL, HI
    SHETH, H
    SPORNS, P
    FOOD RESEARCH INTERNATIONAL, 1992, 25 (05) : 343 - 353
  • [50] Development of monoclonal antibody based antigen capture ELISA to detect chikungunya virus antigen in mosquitoes
    Hundekar, SL
    Thakare, JP
    Gokhale, MD
    Barde, PV
    Argade, SV
    Mourya, DT
    INDIAN JOURNAL OF MEDICAL RESEARCH, 2002, 115 : 144 - 148