Diagnostic system for the detection of severe fever with thrombocytopenia syndrome virus RNA from suspected infected animals

PLoS One. 2021 Jan 28;16(1):e0238671. doi: 10.1371/journal.pone.0238671. eCollection 2021.

Abstract

Background: Severe fever with thrombocytopenia syndrome virus (SFTSV) causes severe hemorrhagic fever in humans and cats. Clinical symptoms of SFTS-infected cats resemble those of SFTS patients, whereas SFTS-contracted cats have high levels of viral RNA loads in the serum and body fluids. Due to the risk of direct infection from SFTS-infected cats to human, it is important to diagnose SFTS-suspected animals. In this study, a reverse transcription polymerase chain reaction (RT-PCR) was newly developed to diagnose SFTS-suspected animals without non-specific reactions.

Methodology/principle findings: Four primer sets were newly designed from consensus sequences constructed from 108 strains of SFTSV. A RT-PCR with these four primer sets successfully and specifically detected four clades of SFTSV. Their limits of detection are 1-10 copies/reaction. Using this RT-PCR, 5 cat cases among 56 SFTS-suspected animal cases were diagnosed as SFTS. From these cats, IgM or IgG against SFTSV were detected by enzyme-linked immunosorbent assay (ELISA), but not neutralizing antibodies by plaque reduction neutralization titer (PRNT) test. This phenomenon is similar to those of fatal SFTS patients.

Conclusion/significance: This newly developed RT-PCR could detect SFTSV RNA of several clades and from SFTS-suspected animals. In addition to ELISA and PRNT test, the useful laboratory diagnosis systems of SFTS-suspected animals has been made in this study.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Viral / immunology
  • Bunyaviridae Infections / virology
  • Cats / virology
  • Diagnostic Tests, Routine / methods
  • Dogs
  • Enzyme-Linked Immunosorbent Assay / methods
  • Female
  • Fever / diagnosis
  • Hemorrhagic Fevers, Viral / diagnosis
  • Hemorrhagic Fevers, Viral / veterinary
  • Hemorrhagic Fevers, Viral / virology
  • Immunoglobulin G / immunology
  • Immunoglobulin M / immunology
  • Japan
  • Male
  • Phlebovirus / genetics*
  • Phlebovirus / metabolism
  • RNA, Viral / blood
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Severe Fever with Thrombocytopenia Syndrome / diagnosis*
  • Severe Fever with Thrombocytopenia Syndrome / veterinary*
  • Severe Fever with Thrombocytopenia Syndrome / virology
  • Thrombocytopenia / diagnosis

Substances

  • Antibodies, Viral
  • Immunoglobulin G
  • Immunoglobulin M
  • RNA, Viral

Grants and funding

The work was partly supported by Japan Agency for Medical Rersearch and Development (AMED) under Grant Number 20fk0108069, 20fk0108081 and Grants-in-Aid for Scientific Research under grant Number 19K06395. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.