Generation of Recombinant Schmallenberg Virus Nucleocapsid Protein in Yeast and Development of Virus-Specific Monoclonal Antibodies

Lazutka, Justas; Žvirblienė, Aurelija; Dalgėdienė, Indrė; Petraitytė-Burneikienė, Rasa; Špakova, Aliona; Sereika, Vilimas; Lelešius, Raimundas; Wernike, Kerstin; Beer, Martin; Sasnauskas, Kęstutis

doi:10.1155/2014/160316

Cited by 8 publications

(9 citation statements)

References 27 publications

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“…The sensitivity of 96 % for serum IgG assay compared to commercial serum IgG ELISA was achieved. This shows a slight improvement over our recently developed indirect serum anti-SBV IgG ELISA [ 16 ], where the sensitivity of the assay was 95 %. A low degree of correlation between optical densities of the same serum samples in commercial and our tests was observed and may suggest that different epitopes on the nucleocapsid protein are recognized in these assays.…”

Section: Discussionmentioning

confidence: 75%

“…The cloning of the SBV N gene and purification under denaturing conditions of the recombinant SBV N antigen was recently described [ 16 ]. Our recent findings indicated that the SBV N protein purified under native conditions exhibited better antigenicity than the SBV N protein purified under denaturing conditions (See Additional file 1 : Figure S1 and Additional file 2 : Figure S2).…”

Section: Methodsmentioning

confidence: 99%

“…Thus, for the development of new diagnostic kits, the SBV N protein purified under native conditions seems to have better antigenic characteristics and sero-diagnostic potential. Saccharomyces cerevisiae AH22-214 yeast transformation with the plasmid vector was described previously [ 16 , 17 ]. In this study the pFGG-SBV-N plasmid vector, containing non-tagged full length SBV N protein coding sequence, was used for transformation.…”

Section: Methodsmentioning

confidence: 99%

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Saliva as an alternative specimen for detection of Schmallenberg virus-specific antibodies in bovines

et al. 2015

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BackgroundSchmallenberg virus (SBV), discovered in continental Europe in late 2011, causes mild clinical signs in adult ruminants, including diarrhoea and reduced milk yield. However, fetal infection can lead to severe malformation in newborn offspring. Enzyme-linked immunosorbent assays (ELISA) are commercially available for detection of SBV-specific antibodies in bovine sera and milk. Here we describe the development and evaluation of an indirect ELISA based on a yeast derived recombinant SBV nucleocapsid protein (N) for the detection of SBV-specific antibodies in bovine saliva. Development of a non-invasive test to detect antibodies in individual bovine saliva samples could potentially provide a test suitable for calves and adult cattle. The aim of this study was to investigate the agreement between the levels of antibodies (IgG) measured in milk and sera, and the level of antibodies (IgG and IgA) in saliva, in comparison with the antibody levels detected in sera and milk with commercially available test.ResultsSerum, milk and saliva samples from 58 cows were collected from three dairy herds in Lithuania and tested for the presence of SBV-specific antibodies. The presence of IgG antibodies was tested in parallel serum and milk samples, while the presence of IgA and IgG antibodies was tested in saliva samples. The presence of SBV-specific IgG and IgA in saliva was tested using an indirect ELISA based on a yeast-derived recombinant N protein. The presence of SBV-specific IgG in milk and sera was tested in parallel using a commercial recombinant protein based test. The sensitivities of the newly developed tests were as follows: 96 % for the IgG serum assay and 94 % for the IgG milk assay and 85 % and 98 % for IgG and IgA in saliva tests, when compared with data generated by a commercial IgG assay.ConclusionsData from testing the saliva IgG and IgA and also the milk and serum IgG with indirect SBV-specific ELISAs showed close agreement with the commercial serum and milk IgG assay data. The level of IgG in saliva was notably lower in comparison to IgA. The newly developed method exhibits the potential to serve as an easily transferable tool for epidemiological studies.Electronic supplementary materialThe online version of this article (doi:10.1186/s12917-015-0552-0) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 75%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Saliva as an alternative specimen for detection of Schmallenberg virus-specific antibodies in bovines

et al. 2015

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“…However, past infections would have been shown by antibody tests. Because bunyaviruses from all vertebrate-infecting genera induce antibodies against the nucleoprotein ( 12 – 14 ), we are confident about our choice of antigen in our assays. Chung et al presented no serologic results to support virus detections ( 9 ).…”

Section: The Studymentioning

confidence: 99%

No Evidence of Gouléako and Herbert Virus Infections in Pigs, Côte d’Ivoire and Ghana

Junglen¹,

Marklewitz²,

Zirkel³

et al. 2015

Emerg. Infect. Dis.

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“…Although the mechanism of overwintering is not known until now and despite a very high seroprevalence of approximately 70% to nearly 100% in ruminants in the initially most affected areas [ 6 , 7 , 8 ], SBV re-appeared in the following vector season [ 9 ]. In 2012, SBV spread across the British Isles to Scotland and Ireland [ 10 , 11 , 12 , 13 , 14 ], reached the Scandinavian countries [ 15 , 16 , 17 ] and Eastern Europe as far as Lithuania [ 18 ], crossed the Alps [ 19 , 20 ], and spread to the Mediterranean area [ 21 , 22 ]. Thus, the virus was detected Europe-wide only 1 year after its initial discovery [ 23 , 24 ].…”

Section: Introductionmentioning

confidence: 99%

Schmallenberg Virus: To Vaccinate, or Not to Vaccinate?

Wernike

Beer

2020

Vaccines

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Schmallenberg virus (SBV), a teratogenic orthobunyavirus that infects predominantly ruminants, emerged in 2011 in Central Europe, spread rapidly throughout the continent, and subsequently established an endemic status with re-circulations to a larger extent every 2 to 3 years. Hence, it represents a constant threat to the continent’s ruminant population when no effective countermeasures are implemented. Here, we discuss potential preventive measures to protect from Schmallenberg disease. Previous experiences with other arboviruses like bluetongue virus have already demonstrated that vaccination of livestock against a vector-transmitted disease can play a major role in reducing or even stopping virus circulation. For SBV, specific inactivated whole-virus vaccines have been developed and marketing authorizations were granted for such preparations. In addition, candidate marker vaccines either as live attenuated, DNA-mediated, subunit or live-vectored preparations have been developed, but none of these DIVA-capable candidate vaccines are currently commercially available. At the moment, the licensed inactivated vaccines are used only to a very limited extent. The high seroprevalence rates induced in years of virus re-occurrence to a larger extent, the wave-like and sometimes hard to predict circulation pattern of SBV, and the expenditures of time and costs for the vaccinations presumably impact on the willingness to vaccinate. However, one should bear in mind that the consequence of seronegative young animals and regular renewed virus circulation might be again more cases of fetal malformation caused by an infection of naïve dams during one of their first gestations. Therefore, an appropriate and cost-effective strategy might be to vaccinate naïve female animals of all affected species before the reproductive age.

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Generation of Recombinant Schmallenberg Virus Nucleocapsid Protein in Yeast and Development of Virus-Specific Monoclonal Antibodies

Cited by 8 publications

References 27 publications

Saliva as an alternative specimen for detection of Schmallenberg virus-specific antibodies in bovines

Saliva as an alternative specimen for detection of Schmallenberg virus-specific antibodies in bovines

No Evidence of Gouléako and Herbert Virus Infections in Pigs, Côte d’Ivoire and Ghana

Schmallenberg Virus: To Vaccinate, or Not to Vaccinate?

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