An International Comparative Analysis of a Competitive ELISA for the Detection of Antibodies to Swine Vesicular Disease Virus

Heckert, Robert A.; Brocchi, Emiliana; Berlinzani, A.; Mackay, D. K. J.

doi:10.1177/104063879801000315

Cited by 4 publications

(8 citation statements)

References 3 publications

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“…The OIE recommended 5B7 cELISA has a standard cut‐off at 70% inhibition (Brocchi et al, ; OIE Manual, ). In this study, a cut‐off of 50% inhibition was used as the negative threshold based on the validation study using the swine sera from Canada (Heckert et al, ). 303 sera without SVDV exposure history identified positive results by the 5B7 cELISA were tested using the SVD‐VLP isotype ELISAs to determine whether the isotype ELISAs could decrease in false‐positive rates.…”

Section: Discussionmentioning

confidence: 99%

“…A panel of SVD reference sera from the Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia‐Romagna (IZSLER) contained 16 positive sera from either outbreak (11 samples from five outbreaks detected in Italy in different years during 1994–2007) or experimentally inoculated pigs (Table ). The true SVD‐positive sera were obtained from pigs experimentally infected with SVDV (Heckert, Brocchi, Berlinzani, & Mackay, ). Serial bleed serum samples were obtained from pigs experimentally infected with SVDV (experiment # 1, 13 pigs, 0 to 14 dpi, 6 of 13 pigs up to 21 dpi) as previously described (Senthilkumaran et al, ) and in a repeated experiment (experiment #2, 6 pigs, 0–28 dpi) using the same experimental procedure.…”

Section: Methodsmentioning

confidence: 99%

“…The 1:15 dilution was chosen, because under this condition, negative sera showed the lowest background and a large range between positive and negative sera. (b) The cut‐off value was set at 50% instead of 70% inhibition based on a previous validation study using the Canadian swine sera (Heckert et al, ). Briefly, microtitre plates were coated 100 μl/well with optimal diluted capture mAb 5B7 (Istituto Zooprofilattico Sperimentale) in a coating buffer (0.06 M Carbonate buffer, pH 9.6) overnight at 4°C.…”

Section: Methodsmentioning

confidence: 99%

“…Results were calculated based on strong positive quality control (Q1) and negative reference sera (Q3) using the following formula: % inhibition = [(Q3 OD – test sample OD)/(Q3 OD – Q1 OD)] × 100%. The cut‐off value was set at 50% inhibition based on a previous validation study using the Canadian swine sera (Heckert et al, ).…”

Section: Methodsmentioning

confidence: 99%

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Development and evaluation of swine vesicular disease isotype‐specific antibody ELISAs based on recombinant virus‐like particles

Yang

Gagliardi

McIntyre

et al. 2019

Transbound Emerg Dis

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Swine vesicular disease (SVD) is a contagious viral disease of pigs. The clinical signs of SVD are indistinguishable from other vesicular diseases, such as senecavirus A infection (SVA) and foot‐and‐mouth disease (FMD). Rapid and accurate diagnostic tests of SVD are considered essential in countries free of vesicular diseases. Competitive ELISA (cELISA) is the serological test used routinely. However, although cELISA is the standard test for SVD antibody testing, this test produces a small number of false‐positive results, which caused problems in international trade. The current project developed a SVD isotype antibody ELISA using recombinant SVD virus‐like particles (VLP) and an SVD‐specific monoclonal antibody (mAb) to reduce the percentage of false positives. The diagnostic specificities of SVD‐VLP isotype ELISAs were 98.7% and 99.6% for IgM and IgG. The SVD isotype ELISAs were SVD‐specific, without cross‐reactivity to other vesicular diseases. A panel of 16 SVD‐positive reference sera was evaluated using the SVD‐VLP isotype ELISAs. All sera were correctly identified as positive by the two combined SVD‐VLP isotype ELISAs. Comparison of the test results showed a high level of correlation between the SVDV antigen isotype ELISAs and SVD‐VLP isotype ELISAs. 303 sera from animals lacking clinical signs and history of SVDV exposure were identified positive using SVD cELISA. These samples were examined using SVD‐VLP isotype ELISAs. Of the 303 serum samples, five were positive for IgM, and five of 303 were positive for IgG. Comparable to virus neutralization test results, SVD isotype ELISAs significantly reduced the false‐positive samples. Based on above test results, the combined use of cELISA and isotype ELISAs can reduce the number of false‐positive samples and the use of time‐consuming virus neutralization tests, with benefit for international trade in swine and related products.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Development and evaluation of swine vesicular disease isotype‐specific antibody ELISAs based on recombinant virus‐like particles

Yang

Gagliardi

McIntyre

et al. 2019

Transbound Emerg Dis

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“…In total, 430 blood samples of wild boars, killed during the 2017-2018 hunting season, were collected in the framework of the Lombardy and Emilia Romagna wildlife monitoring plans for classical swine fever (CSF) and Aujeszky disease and transferred to IZSLER for examination. Serum samples were tested for antibodies against swine vesicular disease virus [30,31], encephalomyocarditis virus [32], glycoprotein E of Aujeszky's disease virus [33], pestivirus (A-D) [34], swine influenza virus type A, subtypes H1N1, H1N2, and H3N2 [35,36], and finally for Brucella spp. (Svanovir Brucella-Ab C-ELISA©).…”

Section: Wild Boarsmentioning

confidence: 99%

Molecular Survey and Phylogenetic Analysis of Atypical Porcine Pestivirus (APPV) Identified in Swine and Wild Boar from Northern Italy

Sozzi¹,

Salogni²,

Lelli³

et al. 2019

Viruses

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Atypical porcine pestivirus (APPV) is a newly recognized member of the Flaviviridae family. This novel porcine pestivirus was first described in 2015 in the USA, where it has been associated with congenital tremor type A-II in new-born piglets. APPV is widely distributed in domestic pigs in Europe and Asia. In this study, a virological survey was performed in Northern Italy to investigate the presence of APPV using molecular methods. Testing of 360 abortion samples from pig herds revealed two APPV strains from distinct provinces in the Lombardy region and testing of 430 wild boar blood samples revealed three strains, one from Lombardy and two from Emilia Romagna. The nucleotide sequencing of a fragment of the nonstructural protein 3-coding region revealed a high similarity to the previously detected European strains (Spanish, German, and Italian) of APPV.

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Swine vesicular disease virus. Pathology of the disease and molecular characteristics of the virion

Escribano-Romero

Jiménez‐Clavero

Ley

2000

Anim. Health. Res. Rev.

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Swine vesicular disease is a highly contagious disease of pigs that is caused by an enterovirus of the family Picornaviridae. The virus is a relatively recent derivative of the human coxsackievirus B5, with which it has high molecular and antigenic homology. The disease is not severe, and affected animals usually show moderate general weakening and slight weight loss that is recovered in few days, as well as vesicular lesions in the mucosa of the mouth and nose and in the interdigital spaces of the feet. However, the similarity of these lesions to those caused by foot-and-mouth disease virus has led to the inclusion of this virus in list A of the Office International des Epizooties. The disease has been eradicated in the European Union except in Italy, where it is considered endemic in the south. Nevertheless, as occasional outbreaks still appear and must be eliminated rapidly, European countries are on the alert and farms are monitored routinely for the presence of the virus. This circumstance has led to a considerable effort to study the pathology of the disease and the molecular biology and antigenicity of the virus, and to the development of optimized methods for the diagnosis of the infection.

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An International Comparative Analysis of a Competitive ELISA for the Detection of Antibodies to Swine Vesicular Disease Virus

Cited by 4 publications

References 3 publications

Development and evaluation of swine vesicular disease isotype‐specific antibody ELISAs based on recombinant virus‐like particles

Development and evaluation of swine vesicular disease isotype‐specific antibody ELISAs based on recombinant virus‐like particles

Molecular Survey and Phylogenetic Analysis of Atypical Porcine Pestivirus (APPV) Identified in Swine and Wild Boar from Northern Italy

Swine vesicular disease virus. Pathology of the disease and molecular characteristics of the virion

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