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Development and evaluation of RT-qPCR assays for two neglected orthobunyaviruses: Oya virus and Ebinur Lake virus.
Liu, Siyuan; Chen, Wei; Nyaruaba, Raphael; Wang, Shunlong; Yang, Cihan; Wu, Qun; Liu, Ying; Liu, Puyu; Wang, Fei; Wang, Jingling; Yuan, Zhiming; Sun, Dingwei; Xia, Han.
Afiliação
  • Liu S; Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, China; University of Chinese Academy of Sciences, Beijing, China.
  • Chen W; Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, China; University of Chinese Academy of Sciences, Beijing, China.
  • Nyaruaba R; Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, China; University of Chinese Academy of Sciences, Beijing, China; Sino-Africa Joint Research Center, Chinese Academy of Sciences, Wuhan, Hubei, China.
  • Wang S; Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, China; University of Chinese Academy of Sciences, Beijing, China.
  • Yang C; Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, China; University of Chinese Academy of Sciences, Beijing, China.
  • Wu Q; Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, China; University of Chinese Academy of Sciences, Beijing, China.
  • Liu Y; Hainan Provincial Center for Disease Control and Prevention, Haikou, China.
  • Liu P; Hainan Provincial Center for Disease Control and Prevention, Haikou, China.
  • Wang F; Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, China.
  • Wang J; Yunnan Tropical and Subtropical Animal Virus Disease Laboratory, Yunnan Animal Science and Veterinary Institute, Kunming, Yunnan Province, China.
  • Yuan Z; Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, China; University of Chinese Academy of Sciences, Beijing, China.
  • Sun D; Hainan Provincial Center for Disease Control and Prevention, Haikou, China. Electronic address: sdw_bmjc@163.com.
  • Xia H; Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, China; University of Chinese Academy of Sciences, Beijing, China; Hubei Jiangxia Laboratory, Wuhan, China. Electronic address: hanxia@wh.iov.cn.
Virus Res ; 339: 199265, 2024 01 02.
Article em En | MEDLINE | ID: mdl-37940076
ABSTRACT

OBJECTIVES:

Oya virus (OYAV) and Ebinur lake virus (EBIV) belong to the genus Orthobunyavirus within the Peribunyaviridae family, and both are recognized as the novel virus with potential threat to the animal or public health. Given their potential to cause outbreaks and their detection in diverse samples across different regions, the need for a reliable and efficient molecular detection method for OYAV and EBIV becomes imperative.

METHODS:

The S-segment of OYAV and EBIV was used for designing specific primer and probe sets, which were employed in a real-time reverse transcription quantitative PCR (RT-qPCR) assay. The analytical performance of these assays, encompassing specificity, sensitivity, reproducibility, and fitness for purpose, was thoroughly evaluated across various sample matrices.

RESULTS:

The developed RT-qPCR assays were very specific to their respective targets. Both assays were highly reproducible (%CV<3) and sensitive with the 95% limit of detection (LOD) of 0.80 PFU/mL for OYAV primer probe set and 0.37 PFU/mL for EBIV primer probe set. Furthermore, the assays fitness for purpose was good as it could detect the specific viruses in virus-spiked serum samples, virus-inoculated mosquito samples, field caught mosquitoes and biting midge samples.

CONCLUSIONS:

Our study has successfully developed specific, sensitive, and reliable RT-qPCR assays for the detection of OYAV and EBIV. These assays hold great promise for their potential application in clinical and field samples in the future.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Orthobunyavirus / Culicidae Limite: Animals Idioma: En Revista: Virus Res Assunto da revista: VIROLOGIA Ano de publicação: 2024 Tipo de documento: Article País de afiliação: China País de publicação: Holanda

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Orthobunyavirus / Culicidae Limite: Animals Idioma: En Revista: Virus Res Assunto da revista: VIROLOGIA Ano de publicação: 2024 Tipo de documento: Article País de afiliação: China País de publicação: Holanda