Development of a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for the detection of porcine pegivirus

Hao Li, Kai Li, Zhen Bi, Jun Gu, Deping Song, Dan Lei, Suoxian Luo, Dongyan Huang, Qiong Wu, Zhen Ding, Leyi Wang, Yu Ye, Yuxin Tang

Research output: Contribution to journalArticlepeer-review

Abstract

A simple and accurate reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay was developed and evaluated for the detection of porcine pegivirus (PPgV). The specific RT-LAMP primers targeting the conserved regions of NS5A genes were designed and used to detect PPgV. The optimal reaction parameter for RT-LAMP assay was 63℃ for 60 min. The detection limit of the RT-LAMP assay was 10 copies of PPgV genome, which was 100 times more sensitive than that of the conventional RT-PCR and comparable to nested RT-PCR and quantitative RT-PCR (qRT-PCR). There was no cross amplification with other related RNA viruses. In the clinical evaluation, the RT-LAMP assay exhibited a similar sensitivity with nested RT-PCR and qRT-PCR. The results indicated that RT-LAMP assay developed in this study could be a highly specific, sensitive, and cost-effective alternative for a rapid detection of PPgV in field settings.

Original languageEnglish (US)
Pages (from-to)59-65
Number of pages7
JournalJournal of Virological Methods
Volume270
DOIs
StatePublished - Aug 2019

Keywords

  • Nested RT-PCR
  • Porcine pegivirus
  • Reverse transcription-loop-mediated isothermal amplification
  • qRT-PCR

ASJC Scopus subject areas

  • Virology

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