Detection of antibodies to avian infectious bronchitis virus by a recombinant nucleocapsid protein-based enzyme-linked immunosorbent assay

Lugovskaya, N. N.; Scherbakov, A. V.; Yakovleva, A. S.; Tsyvanyuk, M.A.; Мудрак, Н. С.; Drygin, V. V.; Борисов, А. В.

doi:10.1016/j.jviromet.2006.03.019

Cited by 19 publications

(23 citation statements)

References 13 publications

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“…As was previously documented, most serological studies including the IDEXX ELISA kit use viral particles of IBV as an antigen for detection of IBV infection. However, preparation of purified virions for use as an ELISA coating antigen is time-consuming and expensive, while recombinant protein expressed in vitro has more advantages as serodiagnostic antigen that it allow better standardization of the tests, reduce the costs of production and purification, and is easy to prepare with large amount (Lugovskaya et al, 2006). In this study, we used GST-fused nsp5 expressed by E. coli as the coating antigen to establish an nsp5-ELISA to detect IBV antibody.…”

Section: Discussionmentioning

confidence: 99%

Development and application of nsp5-ELISA for the detection of antibody to infectious bronchitis virus

Lei

Shi

Sun

et al. 2017

Journal of Virological Methods

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Infectious bronchitis virus (IBV) continues to be one of the most important poultry pathogens worldwide. The current commercially available enzyme-linked immunosorbent assay (ELISA) kits for IBV specific antibody detection are mostly based on the whole virion, and few serological tests based on nonstructural proteins of IBV have been developed. Herein, an alternative indirect ELISA for detection of IBV antibody was developed with IBV nonstructural protein 5 (nsp5) produced by Escherichia coli. Using an indirect immunofluorescence assay (IFA) and a commercial ELISA kit as reference, we optimized the nsp5-ELISA and determined its cut-off as 0.12. The diagnostic sensitivity (DSN), specificity (DSP) and accuracy of the nsp5-ELISA were 93.11%, 95.38% and 93.33%, respectively, compared with IFA in 660 field serum samples, and were 98.11%, 95.00% and 97.62%, respectively, compared with the commercial IBV ELISA kit (IDEXX) in 126 field sera samples. Furthermore, a time course of IBV specific antibody level detected by nsp5-ELISA following IBV infection and vaccination is consistent with that of IBV antibody detected by the commercial ELISA kit. The results presented in this study indicate that nsp5-ELISA has the potential to serve as a rapid, reliable and cost-effective method for IBV antibody detection. This study is the first to report the development of an nsp-based ELISA to detect an antibody to IBV.

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Section: Discussionmentioning

confidence: 99%

Development and application of nsp5-ELISA for the detection of antibody to infectious bronchitis virus

Lei

Shi

Sun

et al. 2017

Journal of Virological Methods

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“…The spike protein, a viral surface glycoprotein, was shown to induce neutralizing antibody response, and the nucleocpasid protein was shown to elicit strong antibody responses [2,5]. Despite the presence and application of IBV vaccines in poultry, there is a high rate of emergence of antigenic variants and recombinant strains, and the lack of cross-protection between different viral genotypes, making disease control difficult and vaccine development rather challenging [15,16]. Therefore, genetic characterization of circulating strains of IBV, appropriate vaccination programs, and application of sensitive diagnostic tests to detect and assess disease risk are important regional, national, and international strategies to control IBV infections [2,8,10,21,23].…”

Section: Introductionmentioning

confidence: 99%

“…Several ELISAs have been developed for detection of antibodies to IBV in chickens. Recombinant antigens used in these ELISAs were either based on the S1 protein [20], which is highly variable, or the N protein and often produced in an E. coli expression system [15]. E. coli is a common flora or pathogen in chickens, thus creating the possibility of serological cross-reactivity and detection of false positives in these diagnostic assays.…”

Section: Introductionmentioning

confidence: 99%

Production of Recombinant N Protein of Infectious Bronchitis Virus Using the Baculovirus Expression System and Its Assessment as a Diagnostic Antigen

Yılmaz

Faburay

Turan

et al. 2018

Appl Biochem Biotechnol

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The avian coronavirus-infectious bronchitis virus (AvCoV-IBV) is recognized as an important avian pathogen, and new viral variants are a continuous threat to the poultry industry worldwide. Sensitive diagnostics and efficacious vaccines are necessary to combat IBV infections in chickens. The aim of this study was to produce recombinant N protein of IBV in the baculovirus system to use in ELISA diagnostic tests in order to enable the assessment of the sero-prevalence and risk of IBV infections in chickens in Turkey. For this, the gene encoding the N protein of the Beaudette strain of IBV was expressed using a recombinant baculovirus expression system. The recombinant N protein was purified using Ni-NTA affinity chromatography. An estimated 50-kDa recombinant protein corresponding to the expected molecular weight of IBV N including the 6xHis tag was detected using an anti-His monoclonal antibody. Specific immunoreactivity of the recombinant protein was confirmed by Western blot using antiserum obtained from vaccinated and naturally infected chicken from Turkey as well as using a monoclonal antibody raised against the N protein of the IBV Massachusetts strain. The results obtained with the in-house ELISA had high agreement with a commercial ELISA. Immunoreactivity analysis using antisera in Western blotting and the in-house ELISA suggests that the recombinant IBV N protein could be broadly cross-reactive with antisera produced against different IBV strains. We conclude that the recombinant baculovirus expressed IBV N protein could serve as a useful diagnostic antigen for detection of IBV infections in chickens by ELISA.

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“…The reaction antigen may be either whole virion or recombinant subunits. In previous works, recombinant spike (Wang et al, 2002) and nucleocapsid protein (Chen et al, 2003;Lugovskaya et al, 2006) expressed in bacteria or insect cells have been used as a coating antigen to improve the efficacy of detection. In addition, an ELISA specific to IBV-IgM was described to detect an early infection (De Wit et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

A type-specific blocking ELISA for the detection of infectious bronchitis virus antibody

Chen

Wang

Cheng

2011

Journal of Virological Methods

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Infectious bronchitis virus (IBV) infection has been a major threat to the poultry industry worldwide. Current commercially available ELISA kits detect group-specific antibodies; however, to understand the status of field infection, a monoclonal antibody (mAb) blocking ELISA (b-ELISA) against local IBVs was developed. The selected mAb showed specificity to Taiwan IBV strains but had no cross-reactivity with the vaccine strain H120. Using the hemagglutination inhibition (HI) test as a standard, the cut-off value, sensitivity, and specificity of a b-ELISA using this mAb were evaluated in 390 field samples. The type-specificity of detection was validated using a panel of chicken hyperimmune sera. The results showed that the b-ELISA demonstrated high sensitivity (98.0%) and specificity (97.2%) of detection. The agreement between the results of the b-ELISA and the HI test was statistically significant (Kappa=0.95), and there was no significant difference between these two methods (McNemar p=0.72). The b-ELISA specifically detected Taiwan IBV serotypes but not three non-Taiwan IBV serotypes nor sera against other avian pathogens. This b-ELISA provides type-specific antibody detection of local IBV strains. It has the potential to serve as a rapid and reliable diagnostic method for IBV clinical infections in the field in Taiwan.

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Detection of antibodies to avian infectious bronchitis virus by a recombinant nucleocapsid protein-based enzyme-linked immunosorbent assay

Cited by 19 publications

References 13 publications

Development and application of nsp5-ELISA for the detection of antibody to infectious bronchitis virus

Development and application of nsp5-ELISA for the detection of antibody to infectious bronchitis virus

Production of Recombinant N Protein of Infectious Bronchitis Virus Using the Baculovirus Expression System and Its Assessment as a Diagnostic Antigen

A type-specific blocking ELISA for the detection of infectious bronchitis virus antibody

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