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Article ; Online: Validation of a Commercial Indirect ELISA Kit for the Detection of

Righi, Cecilia / Iscaro, Carmen / Ferroni, Laura / Rosati, Sergio / Pellegrini, Claudia / Nogarol, Chiara / Rossi, Elisabetta / Dettori, Annalisa / Feliziani, Francesco / Petrini, Stefano

Veterinary sciences

2022 Volume 9, Issue 7

Abstract: In this study, we validated a commercial indirect enzyme-linked immunosorbent assay (ELISA ... of samples (weighted Cohen’s κ: 0.96, 95% CI: 0.78−1.00). The findings demonstrate that the indirect ELISA ...

Abstract	In this study, we validated a commercial indirect enzyme-linked immunosorbent assay (ELISA) to detect antibodies to glycoprotein E (gE) of Bovine alphaherpesvirus 1 (BoHV-1) in bulk milk (BM) samples using the OIE Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. The assay performance characteristics were evaluated using a panel of positive (n = 36) and negative (n = 80) samples with known infectious bovine rhinotracheitis (IBR) status. The assay showed adequate repeatability (within-run and between-run), with a coefficient of variability (CV%) of replicates below 30%; only two 1:40 diluted samples had a CV% above 20%. Additionally, an agreement analysis of the qualitative results of replicates led to a Gwet’s agreement coefficient of 0.99 (95% confidence interval (CI): 0.96−1.00, p < 0.001). The estimated diagnostic sensitivity (DSe) and diagnostic specificity (DSp) were 100% (95% CI: 90.3−100%) and 97.5% (95% CI: 91.3−99.7%), respectively. Overall, a good level of agreement was observed between the assay results and the true IBR status of samples (weighted Cohen’s κ: 0.96, 95% CI: 0.78−1.00). The findings demonstrate that the indirect ELISA kit validated here is an easy-to-use and economical method to differentiate infected and gE-deleted marker vaccine-immunised animals using BM samples.
Language	English
Publishing date	2022-06-22
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2768971-2
ISSN	2306-7381 ; 2306-7381
ISSN (online)	2306-7381
ISSN	2306-7381
DOI	10.3390/vetsci9070311
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Validation of a Commercial Indirect ELISA Kit for the Detection of Bovine alphaherpesvirus 1 (BoHV-1)-Specific Glycoprotein E Antibodies in Bulk Milk Samples of Dairy Cows

Cecilia Righi / Carmen Iscaro / Laura Ferroni / Sergio Rosati / Claudia Pellegrini / Chiara Nogarol / Elisabetta Rossi / Annalisa Dettori / Francesco Feliziani / Stefano Petrini

Veterinary Sciences, Vol 9, Iss 7, p

2022 Volume 311

Abstract: In this study, we validated a commercial indirect enzyme-linked immunosorbent assay (ELISA ... of samples (weighted Cohen’s κ: 0.96, 95% CI: 0.78–1.00). The findings demonstrate that the indirect ELISA ...

Abstract	In this study, we validated a commercial indirect enzyme-linked immunosorbent assay (ELISA) to detect antibodies to glycoprotein E (gE) of Bovine alphaherpesvirus 1 (BoHV-1) in bulk milk (BM) samples using the OIE Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. The assay performance characteristics were evaluated using a panel of positive (n = 36) and negative (n = 80) samples with known infectious bovine rhinotracheitis (IBR) status. The assay showed adequate repeatability (within-run and between-run), with a coefficient of variability (CV%) of replicates below 30%; only two 1:40 diluted samples had a CV% above 20%. Additionally, an agreement analysis of the qualitative results of replicates led to a Gwet’s agreement coefficient of 0.99 (95% confidence interval (CI): 0.96–1.00, p < 0.001). The estimated diagnostic sensitivity (DSe) and diagnostic specificity (DSp) were 100% (95% CI: 90.3–100%) and 97.5% (95% CI: 91.3–99.7%), respectively. Overall, a good level of agreement was observed between the assay results and the true IBR status of samples (weighted Cohen’s κ: 0.96, 95% CI: 0.78–1.00). The findings demonstrate that the indirect ELISA kit validated here is an easy-to-use and economical method to differentiate infected and gE-deleted marker vaccine-immunised animals using BM samples.
Keywords	BoHV-1 ; gE-ELISA ; bulk milk ; kit validation ; Veterinary medicine ; SF600-1100
Subject code	630
Language	English
Publishing date	2022-06-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
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Article: Validation of a Commercial Indirect ELISA Kit for the Detection of Bovine alphaherpesvirus1 (BoHV-1)-Specific Glycoprotein E Antibodies in Bulk Milk Samples of Dairy Cows

Righi, Cecilia / Iscaro, Carmen / Ferroni, Laura / Rosati, Sergio / Pellegrini, Claudia / Nogarol, Chiara / Rossi, Elisabetta / Dettori, Annalisa / Feliziani, Francesco / Petrini, Stefano

Veterinary sciences. 2022 June 22, v. 9, no. 7

2022

Abstract	In this study, we validated a commercial indirect enzyme-linked immunosorbent assay (ELISA) to detect antibodies to glycoprotein E (gE) of Bovine alphaherpesvirus 1 (BoHV-1) in bulk milk (BM) samples using the OIE Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. The assay performance characteristics were evaluated using a panel of positive (n = 36) and negative (n = 80) samples with known infectious bovine rhinotracheitis (IBR) status. The assay showed adequate repeatability (within-run and between-run), with a coefficient of variability (CV%) of replicates below 30%; only two 1:40 diluted samples had a CV% above 20%. Additionally, an agreement analysis of the qualitative results of replicates led to a Gwet’s agreement coefficient of 0.99 (95% confidence interval (CI): 0.96–1.00, p < 0.001). The estimated diagnostic sensitivity (DSe) and diagnostic specificity (DSp) were 100% (95% CI: 90.3–100%) and 97.5% (95% CI: 91.3–99.7%), respectively. Overall, a good level of agreement was observed between the assay results and the true IBR status of samples (weighted Cohen’s κ: 0.96, 95% CI: 0.78–1.00). The findings demonstrate that the indirect ELISA kit validated here is an easy-to-use and economical method to differentiate infected and gE-deleted marker vaccine-immunised animals using BM samples.
Keywords	Bovine alphaherpesvirus 1 ; bulk milk ; confidence interval ; diagnostic sensitivity ; diagnostic specificity ; enzyme-linked immunosorbent assay ; glycoproteins ; infectious bovine rhinotracheitis
Language	English
Dates of publication	2022-0622
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
ZDB-ID	2768971-2
ISSN	2306-7381
ISSN	2306-7381
DOI	10.3390/vetsci9070311
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Field application of an indirect gE ELISA on pooled milk samples for the control of IBR in free and marker vaccinated dairy herds.

Colitti, Barbara / Muratore, Elvira / Careddu, Maria Elena / Bertolotti, Luigi / Iotti, Bryan / Giacobini, Mario / Profiti, Margherita / Nogarol, Chiara / Böttcher, Jens / Ponzo, Andreino / Facelli, Roberto / Rosati, Sergio

BMC veterinary research

2018 Volume 14, Issue 1, Page(s) 387

Abstract: ... to blood serum.: Results: A recently developed indirect gE ELISA showed high versatility when applied ... of IgG from pooled milk samples. The IgG enriched fractions were tested using a gE indirect ELISA ...

Abstract	Background: The aim of the present study was to assess the reliability of a new strategy for monitoring the serological response against Bovine Herpesvirus 1 (BoHV1), the causative agent of infectious bovine rhinotracheitis (IBR). Bulk milk samples have already been identified as cost effective diagnostic matrices for monitoring purposes. Nevertheless, most eradication programs are still based on individual standard assays. In a region of northwestern Italy (Piedmont), the voluntary eradication program for IBR has become economically unsustainable. Being the prevalence of infection still high, glycoprotein E-deleted marker vaccines are commonly used but gE blocking ELISAs are less sensitive on bulk milk samples compared to blood serum. Results: A recently developed indirect gE ELISA showed high versatility when applied to a wide range of matrices. In this study, we applied a faster, cost effective system for the concentration of IgG from pooled milk samples. The IgG enriched fractions were tested using a gE indirect ELISA for monitoring purposes in IBR-positive and IBR-marker-vaccinated herds. Official diagnostic tests were used as gold standard. During a 3 years study, a total 250 herds were involved, including more than 34,500 lactating cows. The proposed method showed a very good agreement with official diagnostic protocols and very good diagnostic performances: only 37 positive animals were not detected across the entire study. Conclusions: The results highlighted the ability of the proposed method to support the surveillance of IBR in the Piedmont region, reducing the costs without affecting the diagnostic performances.
MeSH term(s)	Animals ; Antibodies, Viral/analysis ; Antibodies, Viral/blood ; Cattle ; Dairying/methods ; Enzyme-Linked Immunosorbent Assay/veterinary ; Female ; Herpesvirus 1, Bovine/immunology ; Infectious Bovine Rhinotracheitis/diagnosis ; Infectious Bovine Rhinotracheitis/prevention & control ; Italy ; Milk/chemistry ; Reproducibility of Results ; Vaccination/veterinary
Chemical Substances	Antibodies, Viral
Language	English
Publishing date	2018-12-05
Publishing country	England
Document type	Journal Article
ZDB-ID	2191675-5
ISSN	1746-6148 ; 1746-6148
ISSN (online)	1746-6148
ISSN	1746-6148
DOI	10.1186/s12917-018-1716-5
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Article ; Online: Surveillance of Infectious Bovine Rhinotracheitis in marker-vaccinated dairy herds: Application of a recombinant gE ELISA on bulk milk samples.

Muratore, Elvira / Bertolotti, Luigi / Nogarol, Chiara / Caruso, Claudio / Lucchese, Laura / Iotti, Bryan / Ariello, Dario / Moresco, Angela / Masoero, Loretta / Nardelli, Stefano / Rosati, Sergio

Veterinary immunology and immunopathology

2017 Volume 185, Page(s) 1–6

Abstract: ... methods using the virus neutralization test (VNT) or enzyme-linked immunosorbent assay (ELISA ... methods for the serological diagnosis. Different gE blocking ELISA could be performed to detect ... milk samples, especially in marker-vaccinated herds. A new rec-gE ELISA was recently developed in Italy ...

Abstract	Infectious Bovine Rhinotracheitis (IBR) occurs worldwide, requiring significant resources for eradication programs or surveillance purposes. The status of infection is usually detected by serological methods using the virus neutralization test (VNT) or enzyme-linked immunosorbent assay (ELISA) on individual sera. The gE DIVA (Differentiating Infected from Vaccinated Animals) vaccines approach, adopted in order to reduce the virus circulation and prevent clinical signs, have tightened the range of available methods for the serological diagnosis. Different gE blocking ELISA could be performed to detect specific antibodies in sera of infected or whole virus-vaccinated animals but with less sensitivity if applied to bulk milk samples, especially in marker-vaccinated herds. A new rec-gE ELISA was recently developed in Italy and applied with good performances on blood serum samples. The present paper focuses on the application of a rapid protocol for purification/concentration of immunoglobulin G (IgG) from bulk milk and on the use of the new rec-gE indirect ELISA. The study involved three different partners and 225 herds (12,800 lactating cows) with different official IBR diagnostic statuses. The diagnostic specificity of the method was demonstrated closed to 100% while the diagnostic sensitivity was strictly related to the herd-seroprevalence. Considering 2.5% as the limit of detection of within-herd seropositivity prevalence, the diagnostic sensitivity showed by the proposed method was equal to 100%. A single reactivation of a whole strain vaccine in an old cow was detected inside a group of 67 lactating cows, showing the field applicability of the method.
MeSH term(s)	Animals ; Cattle ; Dairying ; Enzyme-Linked Immunosorbent Assay/methods ; Enzyme-Linked Immunosorbent Assay/veterinary ; Female ; Herpesvirus 1, Bovine/immunology ; Immunoglobulin G/immunology ; Infectious Bovine Rhinotracheitis/diagnosis ; Infectious Bovine Rhinotracheitis/immunology ; Infectious Bovine Rhinotracheitis/prevention & control ; Milk/immunology ; Sensitivity and Specificity ; Viral Proteins/immunology ; Viral Vaccines/immunology
Chemical Substances	Immunoglobulin G ; Viral Proteins ; Viral Vaccines ; bovine herpesvirus type-1 glycoproteins
Language	English
Publishing date	2017-03
Publishing country	Netherlands
Document type	Evaluation Studies ; Journal Article
ZDB-ID	754160-0
ISSN	1873-2534 ; 0165-2427
ISSN (online)	1873-2534
ISSN	0165-2427
DOI	10.1016/j.vetimm.2017.01.003
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Article ; Online: An insight into a combination of ELISA strategies to diagnose small ruminant lentivirus infections.

Veterinary immunology and immunopathology

2013 Volume 152, Issue 3-4, Page(s) 277–288

Abstract: A single broadly reactive standard ELISA is commonly applied to control small ruminant lentivirus ... SRLV) spread, but type specific ELISA strategies are gaining interest in areas with highly prevalent ... among the infected flocks. The ELISA results revealed that none of the assays detected all the infected animals ...

Abstract	A single broadly reactive standard ELISA is commonly applied to control small ruminant lentivirus (SRLV) spread, but type specific ELISA strategies are gaining interest in areas with highly prevalent and heterogeneous SRLV infections. Short (15-residue) synthetic peptides (n=60) were designed in this study using deduced amino acid sequence profiles of SRLV circulating in sheep from North Central Spain and SRLV described previously. The corresponding ELISAs and two standard ELISAs were employed to analyze sera from sheep flocks either controlled or infected with different SRLV genotypes. Two outbreaks, showing SRLV-induced arthritis (genotype B2) and encephalitis (genotype A), were represented among the infected flocks. The ELISA results revealed that none of the assays detected all the infected animals in the global population analyzed, the assay performance varying according to the genetic type of the strain circulating in the area and the test antigen. Five of the six highly reactive (57-62%) single peptide ELISAs were further assessed, revealing that the ELISA based on peptide 98M (type A ENV-SU5, consensus from the neurological outbreak) detected positives in the majority of the type-A specific sera tested (Se: 86%; Sp: 98%) and not in the arthritic type B outbreak. ENV-TM ELISAs based on peptides 126M1 (Se: 82%; Sp: 95%) and 126M2 0,65 0.77 (Se: 68%; Sp: 88%) detected preferentially caprine arthritis encephalitis (CAEV, type B) and visna/maedi (VMV, type A) virus infections respectively, which may help to perform a preliminary CAEV vs. VMV-like typing of the flock. The use of particular peptide ELISAs and standard tests individually or combined may be useful in the different areas under study, to determine disease progression, diagnose/type infection and prevent its spread.
MeSH term(s)	Amino Acid Sequence ; Animals ; Antibodies, Viral/blood ; Antigens, Viral/genetics ; Arthritis-Encephalitis Virus, Caprine/genetics ; Arthritis-Encephalitis Virus, Caprine/immunology ; Disease Outbreaks/veterinary ; Enzyme-Linked Immunosorbent Assay/methods ; Enzyme-Linked Immunosorbent Assay/statistics & numerical data ; Enzyme-Linked Immunosorbent Assay/veterinary ; Genes, gag ; Goats ; Lentivirus Infections/diagnosis ; Lentivirus Infections/epidemiology ; Lentivirus Infections/veterinary ; Molecular Sequence Data ; Phylogeny ; Pneumonia, Progressive Interstitial, of Sheep/diagnosis ; Pneumonia, Progressive Interstitial, of Sheep/epidemiology ; Pneumonia, Progressive Interstitial, of Sheep/immunology ; Sheep ; Sheep Diseases/diagnosis ; Sheep Diseases/epidemiology ; Sheep Diseases/immunology ; Sheep, Domestic ; Spain/epidemiology ; Viral Proteins/genetics ; Viral Proteins/immunology ; Visna/diagnosis ; Visna/epidemiology ; Visna/immunology ; Visna-maedi virus/genetics ; Visna-maedi virus/immunology
Chemical Substances	Antibodies, Viral ; Antigens, Viral ; Viral Proteins
Language	English
Publishing date	2013-04-15
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	754160-0
ISSN	1873-2534 ; 0165-2427
ISSN (online)	1873-2534
ISSN	0165-2427
DOI	10.1016/j.vetimm.2012.12.017
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Article ; Online: Pestivirus infection in cattle dairy farms: E2 glycoprotein ELISA reveals the presence of bovine viral diarrhea virus type 2 in northwestern Italy.

Nogarol, Chiara / Decaro, Nicola / Bertolotti, Luigi / Colitti, Barbara / Iotti, Bryan / Petrini, Stefano / Lucente, Maria Stella / Elia, Gabriella / Perona, Giovanni / Profiti, Margherita / Buonavoglia, Canio / Rosati, Sergio

BMC veterinary research

2017 Volume 13, Issue 1, Page(s) 377

Abstract: ... used in BVDV control programs.: Results: In this study we developed a multiwell antibody ELISA based ...

Abstract	Background: Bovine viral diarrhea virus (BVDV) types 1 and 2 are members of the Pestivirus genus of the Flaviviridae family. This genus also includes the HoBi-like virus, tentatively classified as BVDV type 3. BVDV-1 is widely distributed in Italy despite the extensive use of BVDV-1-based vaccines, while BVDV-2 and HoBi-like Pestivirus have been detected occasionally. Monitoring the occurrence of sporadic or atypical pestiviruses is a useful approach to evaluate the need for additional vaccine strains that can be used in BVDV control programs. Results: In this study we developed a multiwell antibody ELISA based on the recombinant E2 protein of the three bovine pestiviruses. We evaluated the assay's applicability for surveillance purposes using pooled milk samples, each prepared from a maximum of 35 lactating cows and collected from 176 dairy herds. As expected, the majority of the pooled samples reacted to a greater extent against the BVDV-1 E2 antigen. All three milk pools from a single farm reacted to the BVDV-2 antigen, however. Further analysis using spot tests, antigen detection, and sequence analysis of the 5'-UTR region confirmed the presence of five persistently infected calves carrying a BVDV-2a strain. Conclusions: This study highlights for the first time that sporadic circulation of BVDV-2 can be predicted by immunoenzymatic methods in the absence of specific vaccination.
MeSH term(s)	Animals ; Antigens, Viral/immunology ; Bovine Virus Diarrhea-Mucosal Disease/epidemiology ; Bovine Virus Diarrhea-Mucosal Disease/virology ; Cattle ; Diarrhea Virus 2, Bovine Viral/genetics ; Enzyme-Linked Immunosorbent Assay/methods ; Enzyme-Linked Immunosorbent Assay/veterinary ; Italy ; Milk/immunology ; Milk/virology ; Phylogeny ; Recombinant Proteins/immunology
Chemical Substances	Antigens, Viral ; Recombinant Proteins
Language	English
Publishing date	2017-12-04
Publishing country	England
Document type	Journal Article
ISSN	1746-6148
ISSN (online)	1746-6148
DOI	10.1186/s12917-017-1305-z
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Article ; Online: Field application of an indirect gE ELISA on pooled milk samples for the control of IBR in free and marker vaccinated dairy herds

Barbara Colitti / Elvira Muratore / Maria Elena Careddu / Luigi Bertolotti / Bryan Iotti / Mario Giacobini / Margherita Profiti / Chiara Nogarol / Jens Böttcher / Andreino Ponzo / Roberto Facelli / Sergio Rosati

BMC Veterinary Research, Vol 14, Iss 1, Pp 1-

2018 Volume 7

Abstract: ... to blood serum. Results A recently developed indirect gE ELISA showed high versatility when applied to a wide ... of IgG from pooled milk samples. The IgG enriched fractions were tested using a gE indirect ELISA ...

Abstract	Abstract Background The aim of the present study was to assess the reliability of a new strategy for monitoring the serological response against Bovine Herpesvirus 1 (BoHV1), the causative agent of infectious bovine rhinotracheitis (IBR). Bulk milk samples have already been identified as cost effective diagnostic matrices for monitoring purposes. Nevertheless, most eradication programs are still based on individual standard assays. In a region of northwestern Italy (Piedmont), the voluntary eradication program for IBR has become economically unsustainable. Being the prevalence of infection still high, glycoprotein E-deleted marker vaccines are commonly used but gE blocking ELISAs are less sensitive on bulk milk samples compared to blood serum. Results A recently developed indirect gE ELISA showed high versatility when applied to a wide range of matrices. In this study, we applied a faster, cost effective system for the concentration of IgG from pooled milk samples. The IgG enriched fractions were tested using a gE indirect ELISA for monitoring purposes in IBR-positive and IBR-marker-vaccinated herds. Official diagnostic tests were used as gold standard. During a 3 years study, a total 250 herds were involved, including more than 34,500 lactating cows. The proposed method showed a very good agreement with official diagnostic protocols and very good diagnostic performances: only 37 positive animals were not detected across the entire study. Conclusions The results highlighted the ability of the proposed method to support the surveillance of IBR in the Piedmont region, reducing the costs without affecting the diagnostic performances.
Keywords	BoHV1 ; gE ELISA ; Pooled milk ; IBR control ; Veterinary medicine ; SF600-1100
Subject code	630
Language	English
Publishing date	2018-12-01T00:00:00Z
Publisher	BMC
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article: Field application of an indirect gE ELISA on pooled milk samples for the control of IBR in free and marker vaccinated dairy herds

BMC veterinary research. 2018 Dec., v. 14, no. 1

2018

Abstract: ... to blood serum. RESULTS: A recently developed indirect gE ELISA showed high versatility when applied to a wide ... of IgG from pooled milk samples. The IgG enriched fractions were tested using a gE indirect ELISA ...

Abstract	BACKGROUND: The aim of the present study was to assess the reliability of a new strategy for monitoring the serological response against Bovine Herpesvirus 1 (BoHV1), the causative agent of infectious bovine rhinotracheitis (IBR). Bulk milk samples have already been identified as cost effective diagnostic matrices for monitoring purposes. Nevertheless, most eradication programs are still based on individual standard assays. In a region of northwestern Italy (Piedmont), the voluntary eradication program for IBR has become economically unsustainable. Being the prevalence of infection still high, glycoprotein E-deleted marker vaccines are commonly used but gE blocking ELISAs are less sensitive on bulk milk samples compared to blood serum. RESULTS: A recently developed indirect gE ELISA showed high versatility when applied to a wide range of matrices. In this study, we applied a faster, cost effective system for the concentration of IgG from pooled milk samples. The IgG enriched fractions were tested using a gE indirect ELISA for monitoring purposes in IBR-positive and IBR-marker-vaccinated herds. Official diagnostic tests were used as gold standard. During a 3 years study, a total 250 herds were involved, including more than 34,500 lactating cows. The proposed method showed a very good agreement with official diagnostic protocols and very good diagnostic performances: only 37 positive animals were not detected across the entire study. CONCLUSIONS: The results highlighted the ability of the proposed method to support the surveillance of IBR in the Piedmont region, reducing the costs without affecting the diagnostic performances.
Keywords	dairy herds ; Bovine alphaherpesvirus 1 ; diagnostic techniques ; immunoglobulin G ; blood serum ; animals ; monitoring ; bulk milk ; vaccines ; lactation ; veterinary medicine ; enzyme-linked immunosorbent assay ; lactating females ; milk ; infectious bovine rhinotracheitis ; cost effectiveness ; piedmont ; glycoproteins ; Italy
Language	English
Dates of publication	2018-12
Size	p. 387.
Publishing place	BioMed Central
Document type	Article
ISSN	1746-6148
DOI	10.1186/s12917-018-1716-5
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Development and validation of an indirect ELISA as a confirmatory test for surveillance of infectious bovine rhinotracheitis in vaccinated herds.

Bertolotti, Luigi / Muratore, Elvira / Nogarol, Chiara / Caruso, Claudio / Lucchese, Laura / Profiti, Margherita / Anfossi, Laura / Masoero, Loretta / Nardelli, Stefano / Rosati, Sergio

BMC veterinary research

2015 Volume 11, Page(s) 300

Abstract: ... enzyme-linked immunosorbent assay (ELISA) based on BoHV1 glycoprotein E, which was expressed as a secreted recombinant antigen ... in a mammalian cell system. The performance of the new rec-gE ELISA was compared with that of commercially ... 41 and 99.76 %, respectively).: Conclusions: The ELISA performed well, in terms ...

Abstract	Background: Bovine herpesvirus 1 (BoHV1) is a member of the viral subfamily of Alphaherpesvirinae that infects various species, including cattle, sheep, and goats. The virus causes infectious bovine rhinotracheitis (IBR), which is included in a European list of diseases that may require control and eradication programs. The lack of confirmatory tests affects the validity of diagnostic tools, especially those used for vaccinated herds. In this study, we report the development and validation of an indirect enzyme-linked immunosorbent assay (ELISA) based on BoHV1 glycoprotein E, which was expressed as a secreted recombinant antigen in a mammalian cell system. The performance of the new rec-gE ELISA was compared with that of commercially available indirect and/or blocking ELISAs. Results: The sample set included blood sera from animals from IBR-positive farms, IBR-free farms, and marker-vaccinated farms. The indirect ELISA proposed in this study is based on antibody reactivity against BoHV1 gE, and showed high sensitivity and specificity (98.41 and 99.76 %, respectively). Conclusions: The ELISA performed well, in terms of both its diagnostic sensitivity and specificity, and as a confirmatory methodology, and therefore should improve the diagnostic protocols used for IBR surveillance.
MeSH term(s)	Animals ; Antibodies, Viral/blood ; Antigens, Viral/immunology ; Cattle ; Cattle Diseases/prevention & control ; Cell Line ; Enzyme-Linked Immunosorbent Assay/methods ; Herpesvirus 1, Bovine/immunology ; Herpesvirus 1, Bovine/metabolism ; Infectious Bovine Rhinotracheitis/prevention & control ; Population Surveillance ; Recombinant Proteins ; Reproducibility of Results ; Sensitivity and Specificity ; Viral Proteins/immunology ; Viral Vaccines/immunology
Chemical Substances	Antibodies, Viral ; Antigens, Viral ; Recombinant Proteins ; Viral Proteins ; Viral Vaccines ; bovine herpesvirus type-1 glycoproteins
Language	English
Publishing date	2015-12-08
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1746-6148
ISSN (online)	1746-6148
DOI	10.1186/s12917-015-0612-5
Database	MEDical Literature Analysis and Retrieval System OnLINE

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