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Gn protein expressed in plants for diagnosis of severe fever with thrombocytopenia syndrome virus.
Chang, Yu-Chih; Shimoda, Hiroshi; Jiang, Min-Chao; Hsu, Yau-Heiu; Maeda, Ken; Yamada, Yumiko; Hsu, Wei-Li.
Afiliação
  • Chang YC; Doctoral Program in Microbial Genomics, National Chung Hsing University and Academia Sinica, Taichung, Taiwan.
  • Shimoda H; Graduate Institute of Microbiology and Public Health, College of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan.
  • Jiang MC; Laboratory of Veterinary Microbiology, Joint Faculty of Veterinary Medicine, Yamaguchi University, Yamaguchi, Japan.
  • Hsu YH; Doctoral Program in Microbial Genomics, National Chung Hsing University and Academia Sinica, Taichung, Taiwan.
  • Maeda K; Graduate Institute of Biotechnology, National Chung Hsing University, Taichung, Taiwan.
  • Yamada Y; Department of Veterinary Science, National Institute of Infectious Disease, Tokyo, Japan.
  • Hsu WL; Graduate Institute of Microbiology and Public Health, College of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan.
Appl Microbiol Biotechnol ; 108(1): 303, 2024 Apr 19.
Article em En | MEDLINE | ID: mdl-38639795
ABSTRACT
Severe fever with thrombocytopenia syndrome virus (SFTSV) causes the highly fatal disease in humans. To facilitate diagnosis, the native form of subunit glycoprotein (Gn), a prime target for potential vaccines and therapies, was produced in Nicotiana benthamiana using a Bamboo mosaic virus-based vector system. By fusion with secretory signal tags, SSExt, derived from the extension protein, and the (SP)10 motif, the yield of the recombinant Gn (rGn) was remarkably increased to approximately 7 mg/kg infiltrated leaves. Ultimately, an rGn-based ELISA was successfully established for the detection of SFTSV-specific antibodies in serum samples from naturally infected monkeys. As validated with the reference method, the specificity and sensitivity of rGn-ELISA were 94% and 96%, respectively. In conclusion, utilizing well-suited fusion tags facilitates rGn production and purification in substantial quantities while preserving its antigenic properties. The rGn-ELISA, characterized by its commendable sensitivity and specificity could serve as a viable alternative diagnostic method for assessing SFTSV seroprevalence. KEY POINTS • SFTSV Gn, fused with secretory signal tags, was expressed by the BaMV-based vector. • The plant fusion tags increased expression levels and eased the purification of rGn. • The rGn-ELISA was established and validated; its specificity and sensitivity > 94%.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Phlebovirus / Febre Grave com Síndrome de Trombocitopenia Idioma: En Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Taiwan

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Phlebovirus / Febre Grave com Síndrome de Trombocitopenia Idioma: En Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Taiwan