Rapid and sensitive detection of Senecavirus A by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick method

PLoS One. 2019 May 2;14(5):e0216245. doi: 10.1371/journal.pone.0216245. eCollection 2019.

Abstract

Senecavirus A (SVA) is a critical pathogen causing vesicular lesions in sows and acute death of newborn piglets, resulting in very large economic losses in the pig industry. To restrict the transmission of SVA, an establishment of an effective diagnostic method is crucial for the prevention and control of the disease. However, traditional detection methods often have many drawbacks. In this study, reverse transcription loop-mediated isothermal amplification (RT-LAMP) was combined with a lateral flow dipstick (LFD) to detect SVA. The resulting RT-LAMP-LFD assay was performed at 60°C for 50 min and then directly judged on an LFD visualization strip. This method shows high specificity and sensitivity to SVA. The detection limit of RT-LAMP was 4.56x10-8 ng/μL RNA, approximately 11 copies/μL RNA, and it was 10 times more sensitive than RT-PCR. This detection method's positive rate for clinical samples is comparable to that of RT-PCR. This method is time saving and highly efficient and is thus expected to be used to diagnose SVA infections in this field.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Molecular Diagnostic Techniques*
  • Nucleic Acid Amplification Techniques*
  • Picornaviridae / genetics*
  • RNA, Viral / genetics*
  • Reverse Transcription*
  • Sensitivity and Specificity
  • Swine

Substances

  • RNA, Viral

Supplementary concepts

  • Senecavirus A

Grants and funding

This study was supported by 2017 Guangdong Modern Agricultural Pig Industry Technology Promotion System Construction Project (2016LM1103). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.