Development of a real-time loop-mediated isothermal amplification method for the detection of severe fever with thrombocytopenia syndrome virus

J Microbiol. 2020 Aug;58(8):711-715. doi: 10.1007/s12275-020-0109-1. Epub 2020 May 18.

Abstract

Severe fever with thrombocytopenia syndrome (SFTS) is being reported annually in South Korea since its first detection there in 2010. The causal agent is a negative-strand RNA virus 80-100 nm in diameter. It causes fever, thrombocytopenia, leukocytopenia, gastrointestinal symptoms, and neural symptoms. The mortality rate of SFTS was 32.6% among 172 cases reported from 2012 to 2015 in South Korea. Thus, is necessary to develop an effective diagnostic method that selectively identifies the isolates circulating in South Korea. The real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay is a simple, rapid, and sensitive approach for molecular diagnosis. Here, we designed novel primers for this assay and found that the technique had very high specificity, sensitivity, and efficiency. This real-time RT-LAMP approach using the novel primers developed herein can be applied for early diagnosis of SFTSV strains in South Korea to reduce the mortality rate of SFTS.

Keywords: molecular diagnostics; reverse transcription loop-mediated isothermal amplification (LAMP); severe fever with thrombocytopenia syndrome (STFS); virus detection.

MeSH terms

  • DNA Primers / genetics
  • Gastrointestinal Diseases / diagnosis*
  • Gastrointestinal Diseases / virology
  • Humans
  • Molecular Diagnostic Techniques / methods*
  • Nucleic Acid Amplification Techniques / methods*
  • Phlebovirus / genetics*
  • Phlebovirus / isolation & purification
  • RNA, Viral / genetics
  • Republic of Korea
  • Sensitivity and Specificity
  • Severe Fever with Thrombocytopenia Syndrome / diagnosis*
  • Severe Fever with Thrombocytopenia Syndrome / virology

Substances

  • DNA Primers
  • RNA, Viral

Supplementary concepts

  • LAMP assay