scholarly journals Development of a High-Throughput Serum Neutralization Test Using Recombinant Pestiviruses Possessing a Small Reporter Tag

Pathogens ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 188 ◽  
Author(s):  
Madoka Tetsuo ◽  
Keita Matsuno ◽  
Tomokazu Tamura ◽  
Takasuke Fukuhara ◽  
Taksoo Kim ◽  
...  
Keyword(s):  
High Throughput ◽  
Neutralization Test ◽  
Classical Swine Fever ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Border Disease Virus ◽  
Diarrhea Virus ◽  
Serum Neutralization ◽  
Border Disease ◽  
Serum Neutralization Test

A serum neutralization test (SNT) is an essential method for the serological diagnosis of pestivirus infections, including classical swine fever, because of the cross reactivity of antibodies against pestiviruses and the non-quantitative properties of antibodies in an enzyme-linked immunosorbent assay. In conventional SNTs, an immunoperoxidase assay or observation of cytopathic effect after incubation for 3 to 7 days is needed to determine the SNT titer, which requires labor-intensive or time-consuming procedures. Therefore, a new SNT, based on the luciferase system and using classical swine fever virus, bovine viral diarrhea virus, and border disease virus possessing the 11-amino-acid subunit derived from NanoLuc luciferase was developed and evaluated; this approach enabled the rapid and easy determination of the SNT titer using a luminometer. In the new method, SNT titers can be determined tentatively at 2 days post-infection (dpi) and are comparable to those obtained by conventional SNTs at 3 or 4 dpi. In conclusion, the luciferase-based SNT can replace conventional SNTs as a high-throughput antibody test for pestivirus infections.

scholarly journals Serum neutralization as a differential serological test for classical swine fever virus and other pestivirus infections

1999 ◽  
Vol 51 (5) ◽  
pp. 403-408 ◽  
Author(s):  
J.C.M. Paredes ◽  
E.A.S. Oliveira ◽  
L.G. Oliveira ◽  
J.C.A. Rosa ◽  
P.M. Roehe
Keyword(s):  
Classical Swine Fever Virus ◽  
Serological Test ◽  
Sampling Strategy ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Border Disease Virus ◽  
Serum Samples ◽  
Diarrhea Virus ◽  
Serum Neutralization ◽  
Border Disease

Serum neutralization tests (SN) were performed against classical swine fever virus (CSFV), bovine viral diarrhea virus (BVDV) and border disease virus (BDV) on samples of swine serum collected for screening of antibodies to CSFV, in order to determine the SN value as a differential serological test. Ninety-nine sera out of a sample of 16,664 were positive for antibodies to pestiviruses in an ELISA test which did not distinguish antibodies to different pestiviruses. When submitted to SN, 81 sera were positive for CSFV antibodies only. In 17 sera, crossreactive antibodies to either CSFV, BVDV or BDV were detected. In most of these sera (13 out of 17) the differences between SN titres against the three viruses were not sufficient to estimate which was the most likely antibody-inducing virus. It was concluded that, for the SN to be useful in such differentiation, it is essential to examine a sample which must include a representative number of sera from the same farm where suspect animals were detected. When isolated serum samples are examined, such as those obtained with the sampling strategy adopted here, the SN may give rise to inconclusive results.

scholarly journals Identification of a Common Conformational Epitope on the Glycoprotein E2 of Classical Swine Fever Virus and Border Disease Virus

Viruses ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1655
Author(s):  
Yu-Liang Huang ◽  
Denise Meyer ◽  
Alexander Postel ◽  
Kuo-Jung Tsai ◽  
Hsin-Meng Liu ◽  
...  
Keyword(s):  
Disease Virus ◽  
Classical Swine Fever Virus ◽  
Classical Swine Fever ◽  
Conformational Epitope ◽  
Cross Reactivity ◽  
Fever Virus ◽  
Site Directed Mutagenesis ◽  
Border Disease Virus ◽  
Diarrhea Virus ◽  
Border Disease

Classical swine fever virus (CSFV) shares high structural and antigenic homology with bovine viral diarrhea virus (BVDV) and border disease virus (BDV). Because all three viruses can infect swine and elicit cross-reactive antibodies, it is necessary to differentiate among them with regard to serological diagnosis of classical swine fever. To understand the mechanism of cross-reactivity, it is important to define common or specific epitopes of these viruses. For this purpose, epitope mapping of six monoclonal antibodies (mAbs) was performed using recombinant expressed antigenic domains of CSFV and BDV E2 proteins. One CSFV-specific conformational epitope and one CSFV and BDV common epitope within domain B/C of E2 were identified. Site-directed mutagenesis confirmed that residues G725 and V738/I738 of the CSFV-specific epitope and P709/L709 and E713 of the second epitope are important for mAbs binding. Infection of CSFV in porcine cells was significantly reduced after pre-incubation of the cells with the domain B/C of E2 or after pre-incubation of CSFV with the mAbs detecting domain B/C. 3D structural modeling suggested that both epitopes are exposed on the surface of E2. Based on this, the identified epitopes represent a potential target for virus neutralization and might be involved in the early steps of CSFV infection.

scholarly journals Comparative Analysis of Tunisian Sheep-Like Virus, Bungowannah Virus and Border Disease Virus Infection in the Porcine Host

Viruses ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1539
Author(s):  
Denise Meyer ◽  
Alexander Postel ◽  
Anastasia Wiedemann ◽  
Gökce Nur Cagatay ◽  
Sara Ciulli ◽  
...  
Keyword(s):  
Disease Virus ◽  
Classical Swine Fever ◽  
Cross Reactivity ◽  
Border Disease Virus ◽  
Domestic Pigs ◽  
Diarrhea Virus ◽  
Antigenic Relatedness ◽  
Border Disease ◽  
Antibody Assays ◽  
Viral Diarrhea Virus

Apart from the established pestivirus species Pestivirus A to Pestivirus K novel species emerged. Pigs represent not only hosts for porcine pestiviruses, but are also susceptible to bovine viral diarrhea virus, border disease virus (BDV) and other ruminant pestiviruses. The present study focused on the characterization of the ovine Tunisian sheep-like virus (TSV) as well as Bungowannah virus (BuPV) and BDV strain Frijters, which were isolated from pigs. For this purpose, we performed genetic characterization based on complete coding sequences, studies on virus replication in cell culture and in domestic pigs, and cross-neutralization assays using experimentally derived sera. TSV forms a distinct phylogenetic group more closely related to Pestivirus C (classical swine fever virus, CSFV) than to Pestivirus D (BDV). In contrast to BDV and BuPV, TSV replicates by far more efficiently on ovine than on porcine cells. Nevertheless, pigs were susceptible to TSV. As a consequence of close antigenic relatedness of TSV to CSFV, cross-reactivity was detected in CSFV-specific antibody assays. In conclusion, TSV is genetically closely related to CSFV and can replicate in domestic pigs. Due to close antigenic relatedness, field infections of pigs with TSV and other ruminant pestiviruses can interfere with serological diagnosis of classical swine fever.

scholarly journals Pestivirus infections in cervids from the Czech Republic

2009 ◽  
Vol 54 (No. 4) ◽  
pp. 191-193
Author(s):  
K. Sedlak ◽  
T. Girma ◽  
J. Holejsovsky
Keyword(s):  
Czech Republic ◽  
Cervus Elaphus ◽  
Red Deer ◽  
Competitive Inhibition ◽  
Enzyme Linked Immunosorbent Assay ◽  
Border Disease Virus ◽  
Serum Samples ◽  
The Czech Republic ◽  
Diarrhea Virus ◽  
Border Disease

372 sera of cervids from the Czech Republic were examined for antibodies to the bovine viral diarrhea virus (BVDV) and border disease virus (BDV) by competitive-inhibition enzyme-linked immunosorbent assay (ELISA), and for the presence of the BVDV by AgELISA. Antibodies to BVDV/BDV were found in 0.6% (two positive/305 tested) red deer (<I>Cervus elaphus</I>). BVDV/BDV antibodies were not found in four sika deer (<I>Cervus Nippon</I>) and 63 fallow deer (<I>Dama dama</I>). All serum samples were BVDV antigen negative. Our results confirmed that red deer in the Czech Republic are only rarely infected with Pestiviruses. This was the first survey of pestiviruses in farmed and wild cervids in the Czech Republic.

scholarly journals The development and application of an indirect Elisa test for the detection of antibodies to bovine respiratory syncytial virus in blood serum 

2001 ◽  
Vol 46 (No. 2) ◽  
pp. 29-34 ◽  
Author(s):  
K. Kovařčík
Keyword(s):  
Respiratory Syncytial Virus ◽  
Neutralization Test ◽  
Elisa Test ◽  
Bovine Respiratory Syncytial Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Neutralization ◽  
Specificity And Sensitivity ◽  
Reference Serum ◽  
Serum Neutralization Test ◽  
Syncytial Virus

We developed an indirect enzyme-linked immunosorbent assay (ELISA) for detection of serum antibodies to bovine respiratory syncytial virus. For evaluation of the newly developed ELISA, field sera collected from 549 head of cattle in the Czech Republic were tested in parallel by a serum neutralization test. The tests showed 98.36% agreement. The specificity and sensitivity of the ELISA relative to serum neutralization test was 97.00% (226/233) and 99.37% (314/316), respectively. Tissue culture-grown viral antigen was used in the tests. The corrected optical density (COD) of each sample tested at dilution 1/100 was expressed as a percentage of the COD of a positive reference serum included on each plate, this value was the sample/positive (S/P) ratio. We determined the relationship between the S/P ratio (%) obtained at a dilution 1/100 and the end point titer calculated by serum neutralization test (r = 0.9743). The ELISA test was evaluated by testing acute and convalescent (3 wk later) serum pairs from 9 head of cattle with confirmed BRSV infection for demonstration of seroconversion. The ELISA test demonstrated a clear increase of the S/P ratio (%) between acute and convalescent serum pairs (on average 42.2 &plusmn; 13.1).

scholarly journals Comparison of the Serum Neutralization Test and a Competitive Enzyme-Linked Immunosorbent Assay for the Detection of Antibodies to Vesicular Stomatitis Virus New Jersey and Vesicular Stomatitis Virus Indiana

2002 ◽  
Vol 14 (3) ◽  
pp. 240-242 ◽  
Author(s):  
Juan Francisco Alvarado ◽  
Gaby Dolz ◽  
Marco V. Herrero ◽  
Brian McCluskey ◽  
Mo Salman
Keyword(s):  
New Jersey ◽  
Confidence Interval ◽  
Vesicular Stomatitis Virus ◽  
Immunosorbent Assay ◽  
Neutralization Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Serum Neutralization ◽  
Serum Neutralization Test ◽  
Vesicular Stomatitis

A competitive enzyme-linked immunosorbent assay (C-ELISA) for the detection of antibodies against vesicular stomatitis virus New Jersey (VSV-NJ) and vesicular stomatitis virus Indiana (VSV-IN) was compared with the serum neutralization test (SNT) using 1,106 serum samples obtained from dairy cattle on sentinel study farms in the Poás region of Costa Rica. Kappa coefficients between the C-ELISA and the SNT were 0.8871 (95% confidence interval [CI]: 0.8587–0.9155) and 0.6912 (95% CI: 0.6246–0.7577) for the VSV-NJ and VSV-IN tests, respectively. These results indicate good to excellent agreement between the 2 tests under these conditions.

scholarly journals Pestivirus Infections in Semi-Domesticated Eurasian Tundra Reindeer (Rangifer tarandus tarandus): A Retrospective Cross-Sectional Serological Study in Finnmark County, Norway

Viruses ◽  
2019 ◽  
Vol 12 (1) ◽  
pp. 29 ◽  
Author(s):  
Carlos G. das Neves ◽  
Jonas Johansson Wensman ◽  
Ingebjørg Helena Nymo ◽  
Eystein Skjerve ◽  
Stefan Alenius ◽  
...  
Keyword(s):  
Rangifer Tarandus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Border Disease Virus ◽  
Serum Samples ◽  
Cross Sectional ◽  
Diarrhea Virus ◽  
Border Disease ◽  
Serological Studies ◽  
Viral Diarrhea Virus

Members of the Pestivirus genus (family Flaviviridae) cause severe and economically important diseases in livestock. Serological studies have revealed the presence of pestiviruses in different cervid species, including wild and semi-domesticated Eurasian tundra reindeer. In this retrospective study, serum samples collected between 2006 and 2008 from 3339 semi-domesticated Eurasian reindeer from Finnmark County, Norway, were tested for anti-pestivirus antibodies using an enzyme linked immunosorbent assay (ELISA) and a subset of these by virus neutralization test (VNT). A seroprevalence of 12.5% was found, varying from 0% to 45% among different herding districts, and 20% in western Finnmark, as compared to 1.7% in eastern Finnmark. Seroprevalence increased with age. Pestivirus-specific RNA was not detected in any of the 225 serum samples tested by real-time RT-PCR. Based on VNT results, using a panel of one bovine viral diarrhea virus (BVDV) strain and two border disease virus (BDV) strains, the virus is most likely a reindeer-specific pestivirus closely related to BDV. A characterization of the causative virus and its pathogenic impact on reindeer populations, as well as its potential to infect other domestic and wild ruminants, should be further investigated.

scholarly journals Influence of NS5A protein of classical swine fever virus (CSFV) on CSFV internal ribosome entry site-dependent translation

2009 ◽  
Vol 90 (12) ◽  
pp. 2923-2928 ◽  
Author(s):  
Ming Xiao ◽  
Yujing Wang ◽  
Zailing Zhu ◽  
Jialin Yu ◽  
Lingzhu Wan ◽  
...  
Keyword(s):  
Classical Swine Fever Virus ◽  
Internal Ribosome Entry Site ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Border Disease Virus ◽  
Dependent Manner ◽  
Entry Site ◽  
Internal Ribosome Entry ◽  
Diarrhea Virus ◽  
Border Disease

An internal ribosome entry site (IRES) present in the 5′ untranslated region (UTR) promotes translation of classical swine fever virus (CSFV) genomes. Using an in vitro system with monocistronic reporter RNA containing the CSFV 5′UTR, this study found that CSFV NS5A decreased CSFV IRES-mediated translation in a dose-dependent manner. Deletion analysis showed that the region responsible for repressing CSFV IRES activity might cover aa  390–414, located in the C-terminal half of CSFV NS5A. Triple and single alanine-scanning mutagenesis revealed that the inhibitory effect on CSFV IRES-directed translation mapped to the K399, T401, E406 and L413 residues of NS5A. These important amino acids were also found to be present in the NS5A proteins of bovine viral diarrhea virus (BVDV)-1, BVDV-2, border disease virus and hepatitis C virus, indicating that NS5A may play an important role in the switch from translation to replication in these viruses.

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