Detection of antibodies against African swine fever virus using infected monolayers and monoclonal antibodies

1988 ◽  
Vol 122 (22) ◽  
pp. 536-539 ◽  
Author(s):  
G. Wensvoort ◽  
C. Terpstra ◽  
M. Bloemraad
Keyword(s):  
Monoclonal Antibodies ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus

scholarly journals Establishment of a Blocking ELISA Detection Method for Against African Swine Fever Virus p30 Antibody

2021 ◽  
Vol 8 ◽  
Author(s):  
Xuexiang Yu ◽  
Xianjing Zhu ◽  
Xiaoyu Chen ◽  
Dongfan Li ◽  
Qian Xu ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Confidence Interval ◽  
Roc Analysis ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Diagnostic Sensitivity ◽  
Cutoff Value ◽  
Blocking Elisa

African swine fever (ASF) is a highly lethal hemorrhagic viral disease of domestic pigs caused by African swine fever virus (ASFV). A sensitive and reliable serological diagnostic assay is required, so laboratories can effectively and quickly detect ASFV infection. The p30 protein is abundantly expressed early in cells and has excellent antigenicity. Therefore, this study aimed to produce and characterize p30 monoclonal antibodies with an ultimate goal of developing a monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) for ASFV antibody detection. Three monoclonal antibodies against p30 protein that were expressed in E. coli were generated, and their characterizations were investigated. Furthermore, a blocking ELISA based on a monoclonal antibody was developed. To evaluate the performance of the assay, 186 sera samples (88 negative and 98 positive samples) were analyzed and a receiver-operating characteristic (ROC) analysis was applied to determine the cutoff value. Based on the ROC analysis, the area under the curve (AUC) was 0.997 (95% confidence interval: 99.2 to 100%). Besides, a diagnostic sensitivity of 97.96% (95% confidence interval: 92.82 to 99.75%) and a specificity of 98.96% (95% confidence interval: 93.83 to 99.97%) were achieved when the cutoff value was set to 38.38%. Moreover, the coefficients of inter- and intra-batches were <10%, indicating the good repeatability of the method. The maximum dilution of positive standard serum detected by this ELISA method was 1:512. The blocking ELISA was able to detect seroconversion in two out of five pigs at 10 Dpi and the p30 response increasing trend through the time course of the study (0–20 Dpi). In conclusion, the p30 mAb-based blocking ELISA developed in this study demonstrated a high repeatability with maximized diagnostic sensitivity and specificity. The assay could be a useful tool for field surveillance and epidemiological studies in swine herd.

Development and characterization of monoclonal antibodies against p30 protein of African swine fever virus

2019 ◽  
Vol 269 ◽  
pp. 197632 ◽  
Author(s):  
Vlad Petrovan ◽  
Fangfeng Yuan ◽  
Yanhua Li ◽  
Pengcheng Shang ◽  
Maria V. Murgia ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus

Monoclonal antibodies specific for African swine fever virus proteins.

1985 ◽  
Vol 54 (1) ◽  
pp. 199-206 ◽  
Author(s):  
A Sanz ◽  
B García-Barreno ◽  
M L Nogal ◽  
E Viñuela ◽  
L Enjuanes
Keyword(s):  
Monoclonal Antibodies ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Virus Proteins

A colloidal gold test strip assay for the detection of African swine fever virus based on two monoclonal antibodies against P30

2021 ◽  
Vol 166 (3) ◽  
pp. 871-879
Author(s):  
Xinyu Zhang ◽  
Xiaoyu Liu ◽  
Xiaodong Wu ◽  
Weijie Ren ◽  
Yanli Zou ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Colloidal Gold ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Test Strip ◽  
Fever Virus

A comparison of two indirect immune electron microscopy procedures using monoclonal antibody to localize specific African swine fever virus (ASFV) proteins in infected cells and virus

1983 ◽  
Vol 41 ◽  
pp. 800-801
Author(s):  
G. J. Letchworth III ◽  
T. C. Whyard ◽  
S. H. Wool
Keyword(s):  
Electron Microscopy ◽  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Grid Method ◽  
Viral Proteins ◽  
Fever Virus ◽  
Immune Electron Microscopy ◽  
Infected Cells

The primary purpose of this study was to devise a procedure in which a large number of monoclonal antibodies could be tested against several African swine fever virus (ASFV) preparations (virus, infected cells). The procedure had to be rapid, use a small sample and provide for some TEM.Monoclonal antibodies were produced by standard methods using myeloma cells and lymphocytes from mice immunized against ASF viral proteins. Antibodies were characterized by immunoprecipitation of radio-labelled viral proteins (Fig 1).The viral preparations used in this study were either infected macrophages, culture fluid supernatant from ASFV-infected macrophages or red blood cells from infected pigs. Each of the three viral preparations was tested by two indirect immune electron microscopy (IEM) methods; a grid method and a test tube method. In the grid method, formvar coated grids were floated on a viral preparation then for one hour each on the mouse monoclonal antibody and finally on ferritin tagged sheep anti-mouse IgG with washes between each step.

scholarly journals Linear epitopes in African swine fever virus p72 recognized by monoclonal antibodies prepared against baculovirus-expressed antigen

2018 ◽  
Vol 30 (3) ◽  
pp. 406-412 ◽  
Author(s):  
Mallory E. Heimerman ◽  
Maria V. Murgia ◽  
Ping Wu ◽  
Andre D. Lowe ◽  
Wei Jia ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Major Capsid Protein ◽  
Vaccine Development ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Vero Cells ◽  
Fever Virus ◽  
E Coli ◽  
Important Target ◽  
Linear Epitopes

Protein p72 is the major capsid protein of African swine fever virus (ASFV) and is an important target for test and vaccine development. Monoclonal antibodies (mAbs) were prepared against a recombinant antigenic fragment, from amino acid (aa) 20–303, expressed in baculovirus. A total of 29 mAbs were recovered and tested by immunofluorescent antibody (IFA) staining on ASFV Lisbon-infected Vero cells. Six antibodies were IFA-positive and selected for further characterization. Epitope mapping was performed against overlapping polypeptides expressed in E. coli and oligopeptides. Based on oligopeptide recognition, the mAbs were divided into 4 groups: mAb 85 (aa 165–171); mAbs 65-3 and 6H9-1 (aa 265–280); mAbs 8F7-3 and 23 (aa 280–294); and mAb 4A4 (aa 290–303). All mAbs were located within a highly conserved region in p72. This panel of antibodies provides the opportunity to develop new assays for the detection of ASFV antibody and antigen.

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