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


Journal

PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081

Informations de publication

Date de publication:
2021
Historique:
received: 03 09 2020
accepted: 04 12 2020
entrez: 28 1 2021
pubmed: 29 1 2021
medline: 1 5 2021
Statut: epublish

Résumé

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. 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. 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.

Sections du résumé

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.

Identifiants

pubmed: 33507990
doi: 10.1371/journal.pone.0238671
pii: PONE-D-20-25871
pmc: PMC7842937
doi:

Substances chimiques

Antibodies, Viral 0
Immunoglobulin G 0
Immunoglobulin M 0
RNA, Viral 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

e0238671

Déclaration de conflit d'intérêts

The authors have declared that no competing interests exist.

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Auteurs

Eun-Sil Park (ES)

Department of Veterinary Science, National Institute of Infectious Diseases, Shinjuku, Tokyo, Japan.

Osamu Fujita (O)

Department of Veterinary Science, National Institute of Infectious Diseases, Shinjuku, Tokyo, Japan.

Masanobu Kimura (M)

Department of Veterinary Science, National Institute of Infectious Diseases, Shinjuku, Tokyo, Japan.

Akitoyo Hotta (A)

Department of Veterinary Science, National Institute of Infectious Diseases, Shinjuku, Tokyo, Japan.

Koichi Imaoka (K)

Department of Veterinary Science, National Institute of Infectious Diseases, Shinjuku, Tokyo, Japan.

Masayuki Shimojima (M)

Department of Virology I, National Institute of Infectious Diseases, Shinjuku, Tokyo, Japan.

Masayuki Saijo (M)

Department of Virology I, National Institute of Infectious Diseases, Shinjuku, Tokyo, Japan.

Ken Maeda (K)

Department of Veterinary Science, National Institute of Infectious Diseases, Shinjuku, Tokyo, Japan.
Laboratory of Veterinary Microbiology, Yamaguchi University, Yamaguchi, Japan.

Shigeru Morikawa (S)

Department of Veterinary Science, National Institute of Infectious Diseases, Shinjuku, Tokyo, Japan.
Department of Microbiology, Faculty of Veterinary Medicine, Okayama University of Science, Imabari, Ehime, Japan.

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Classifications MeSH