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Development of a recombinant protein-based ELISA for detection of antibodies against bovine herpesvirus 6 (BoHV6)

Piotr Kubiś

Piotr Kubiś

Department of Biochemistry, National Veterinary Research InstitutePuławy, Poland
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Kubiś, Piotr
 e 
Jacek Kuźmak

Jacek Kuźmak

Department of Biochemistry, National Veterinary Research InstitutePuławy, Poland
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Kuźmak, Jacek
  
19 dic 2023
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Pubblicato online: 19 dic 2023
Pagine: 509 - 515
Ricevuto: 08 ott 2023
Accettato: 05 dic 2023
DOI: https://doi.org/10.2478/jvetres-2023-0069
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© 2023 Piotr Kubiś et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Introduction

Bovine herpesvirus 6 (BoHV6) belongs to the Herpesviridae family, Gammaherpesvirinae subfamily and Macavirus genus. It is common in cattle, but was also detected in American bison (Bison bison) and water buffalo (Bubalus bubalis). The aim of the experiment was to develop an ELISA for serological examination of cattle sera for the presence of anti-BoHV6 specific antibodies.

Material and Methods

Viral DNA from a BoHV6-positive cow was amplified by qPCR and the resulting fragments of the gB and gH genes encoding glycoproteins B and H (gB and gH) were cloned into the pLATE52 vector to express recombinant gB (rgB) and gH (rgH) in Rosetta (DE3) E. coli. The expressed recombinant proteins were used as antigens in the developed ELISA.

Results

The proteins expressed had the expected molecular weight. A total of 143 sera were examined, and 141 of them were positive, according to the chosen cut-off values of 9% and 10% for the sample-to-positive ratios of the rgB and rgH antigens, respectively.

Conclusion

The rgB and rgH recombinant antigens of BoHV6 were successfully expressed in E. coli and successfully used in a newly developed ELISA.
Lingua:
Inglese
Frequenza di pubblicazione:
4 volte all'anno
Argomenti della rivista:
Scienze biologiche, Biologia molecolare, Microbiologia e virologia, Scienze della vita, altro, Medicina, Medicina veterinaria
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