Diagnostic system for the detection of severe fever with thrombocytopenia syndrome virus RNA from suspected infected animals
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 dir...
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Published in | PloS one Vol. 16; no. 1; p. e0238671 |
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Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English |
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28.01.2021
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Abstract | 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. |
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AbstractList | 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. BackgroundSevere 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 findingsFour 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/significanceThis 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. 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.BACKGROUNDSevere 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.METHODOLOGY/PRINCIPLE FINDINGSFour 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.CONCLUSION/SIGNIFICANCEThis 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. 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. 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. |
Audience | Academic |
Author | Fujita, Osamu Morikawa, Shigeru Shimojima, Masayuki Saijo, Masayuki Park, Eun-sil Hotta, Akitoyo Kimura, Masanobu Imaoka, Koichi Maeda, Ken |
AuthorAffiliation | 3 Laboratory of Veterinary Microbiology, Yamaguchi University, Yamaguchi, Japan 2 Department of Virology I, National Institute of Infectious Diseases, Shinjuku, Tokyo, Japan University of Texas Medical Branch at Galveston, UNITED STATES 4 Department of Microbiology, Faculty of Veterinary Medicine, Okayama University of Science, Imabari, Ehime, Japan 1 Department of Veterinary Science, National Institute of Infectious Diseases, Shinjuku, Tokyo, Japan |
AuthorAffiliation_xml | – name: University of Texas Medical Branch at Galveston, UNITED STATES – name: 4 Department of Microbiology, Faculty of Veterinary Medicine, Okayama University of Science, Imabari, Ehime, Japan – name: 3 Laboratory of Veterinary Microbiology, Yamaguchi University, Yamaguchi, Japan – name: 2 Department of Virology I, National Institute of Infectious Diseases, Shinjuku, Tokyo, Japan – name: 1 Department of Veterinary Science, National Institute of Infectious Diseases, Shinjuku, Tokyo, Japan |
Author_xml | – sequence: 1 givenname: Eun-sil surname: Park fullname: Park, Eun-sil – sequence: 2 givenname: Osamu surname: Fujita fullname: Fujita, Osamu – sequence: 3 givenname: Masanobu surname: Kimura fullname: Kimura, Masanobu – sequence: 4 givenname: Akitoyo surname: Hotta fullname: Hotta, Akitoyo – sequence: 5 givenname: Koichi surname: Imaoka fullname: Imaoka, Koichi – sequence: 6 givenname: Masayuki surname: Shimojima fullname: Shimojima, Masayuki – sequence: 7 givenname: Masayuki orcidid: 0000-0001-5458-7298 surname: Saijo fullname: Saijo, Masayuki – sequence: 8 givenname: Ken surname: Maeda fullname: Maeda, Ken – sequence: 9 givenname: Shigeru orcidid: 0000-0001-6249-6716 surname: Morikawa fullname: Morikawa, Shigeru |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/33507990$$D View this record in MEDLINE/PubMed |
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CitedBy_id | crossref_primary_10_1292_jvms_22_0236 crossref_primary_10_1016_j_trac_2021_116460 crossref_primary_10_3390_v14091963 crossref_primary_10_3346_jkms_2025_40_e69 crossref_primary_10_1292_jvms_22_0255 crossref_primary_10_7883_yoken_JJID_2021_796 crossref_primary_10_3390_ani11061754 crossref_primary_10_3390_v13040693 crossref_primary_10_1292_jvms_21_0519 |
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References_xml | – volume: 24 start-page: 802 issue: 10 year: 2018 ident: pone.0238671.ref019 article-title: Seroprevalence of severe fever with thrombocytopenia syndrome (SFTS) virus antibodies in humans and animals in Ehime prefecture, Japan, an endemic region of SFTS publication-title: Journal of Infection and Chemotherapy doi: 10.1016/j.jiac.2018.06.007 – volume: 364 start-page: 1523 issue: 16 year: 2011 ident: pone.0238671.ref001 article-title: Fever with thrombocytopenia associated with a novel bunyavirus in China publication-title: N Engl J Med doi: 10.1056/NEJMoa1010095 – volume: 209 start-page: 816 issue: 6 year: 2013 ident: pone.0238671.ref002 article-title: The First Identification and Retrospective Study of Severe Fever With Thrombocytopenia Syndrome in Japan publication-title: The Journal of Infectious Diseases doi: 10.1093/infdis/jit603 – volume: 9 start-page: 11990 issue: 1 year: 2019 ident: pone.0238671.ref014 article-title: Severe Fever with Thrombocytopenia Syndrome Phlebovirus causes lethal viral hemorrhagic fever in cats publication-title: Sci Rep doi: 10.1038/s41598-019-48317-8 – volume: 8 start-page: 9 issue: 1 year: 2017 ident: pone.0238671.ref012 article-title: Molecular detection of severe fever with thrombocytopenia syndrome virus (SFTSV) in feral cats from Seoul, Korea publication-title: Ticks Tick Borne Dis doi: 10.1016/j.ttbdis.2016.08.005 – volume: 19 start-page: 756 issue: 5 year: 2013 ident: pone.0238671.ref008 article-title: Severe fever with thrombocytopenia syndrome virus among domesticated animals, China publication-title: Emerg Infect Dis – volume: 24 start-page: 1726 issue: 9 year: 2018 ident: pone.0238671.ref013 article-title: Fatal Tickborne Phlebovirus Infection in Captive Cheetahs, Japan publication-title: Emerg Infect Dis doi: 10.3201/eid2409.171667 – volume: 25 start-page: 1029 issue: 5 year: 2019 ident: pone.0238671.ref004 article-title: Endemic Severe Fever with Thrombocytopenia Syndrome, Vietnam publication-title: Emerging Infectious Disease journal doi: 10.3201/eid2505.181463 – volume: 52 start-page: 3325 issue: 9 year: 2014 ident: pone.0238671.ref015 article-title: Sensitive and Specific PCR Systems for Detection of Both Chinese and Japanese Severe Fever with Thrombocytopenia Syndrome Virus Strains and Prediction of Patient Survival Based on Viral Load publication-title: Journal of Clinical Microbiology doi: 10.1128/JCM.00742-14 – volume: 33 start-page: 1870 issue: 7 year: 2016 ident: pone.0238671.ref016 article-title: MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets publication-title: Mol Biol Evol doi: 10.1093/molbev/msw054 – volume: 95 start-page: e2533 issue: 4 year: 2016 ident: pone.0238671.ref009 article-title: Natural Transmission Model for Severe Fever With Thrombocytopenia Syndrome Bunyavirus in Villages of Hubei Province, China publication-title: Medicine (Baltimore) doi: 10.1097/MD.0000000000002533 – volume: 67 start-page: 1 issue: 1 year: 2014 ident: pone.0238671.ref011 article-title: A cross-sectional survey of severe fever with thrombocytopenia syndrome virus infection of domestic animals in Laizhou City, Shandong Province, China publication-title: Jpn J Infect Dis doi: 10.7883/yoken.67.1 – volume: 9 start-page: 148 issue: 1 year: 2020 ident: pone.0238671.ref005 article-title: The first discovery of severe fever with thrombocytopenia syndrome virus in Taiwan publication-title: Emerging Microbes & Infections doi: 10.1080/22221751.2019.1710436 – volume: 41 start-page: 95 year: 1999 ident: pone.0238671.ref017 article-title: BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT publication-title: Nucleic Acids Symposium Series – volume: 212 start-page: 889 issue: 6 year: 2015 ident: pone.0238671.ref006 article-title: Phylogenetic and Geographic Relationships of Severe Fever With Thrombocytopenia Syndrome Virus in China, South Korea, and Japan publication-title: J Infect Dis doi: 10.1093/infdis/jiv144 – volume: 19 start-page: 1892 issue: 11 year: 2013 ident: pone.0238671.ref003 article-title: Severe fever with thrombocytopenia syndrome, South Korea, 2012 publication-title: Emerg Infect Dis – volume: 13 start-page: 1471 issue: 134 year: 2012 ident: pone.0238671.ref018 article-title: Primer-BLAST: A tool to design target-specific primers for polymerase chain reaction publication-title: BMC Bioinformatics – volume: 8 start-page: 626 issue: 4 year: 2017 ident: pone.0238671.ref010 article-title: Molecular detection and phylogenetic analysis of severe fever with thrombocytopenia syndrome virus in shelter dogs and cats in the Republic of Korea publication-title: Ticks Tick Borne Dis doi: 10.1016/j.ttbdis.2017.04.008 – volume: 24 start-page: 90 issue: 2 year: 2014 ident: pone.0238671.ref007 article-title: Systematic review of severe fever with thrombocytopenia syndrome: virology, epidemiology, and clinical characteristics publication-title: Rev Med Virol doi: 10.1002/rmv.1776 |
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Snippet | Severe fever with thrombocytopenia syndrome virus (SFTSV) causes severe hemorrhagic fever in humans and cats. Clinical symptoms of SFTS-infected cats resemble... Background Severe fever with thrombocytopenia syndrome virus (SFTSV) causes severe hemorrhagic fever in humans and cats. Clinical symptoms of SFTS-infected... BackgroundSevere fever with thrombocytopenia syndrome virus (SFTSV) causes severe hemorrhagic fever in humans and cats. Clinical symptoms of SFTS-infected cats... |
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SubjectTerms | Animals Antibodies Antibodies, Viral - immunology Antigens Biology and Life Sciences Bunyaviridae Infections - virology Cats - virology Computer and Information Sciences Deoxyribonucleic acid Diagnosis Diagnostic systems Diagnostic Tests, Routine - methods DNA DNA polymerase Dogs Domestic animals Editing Enzyme-Linked Immunosorbent Assay - methods Female Fever Fever - diagnosis Funding Genetic aspects Hemorrhage Hemorrhagic fever Hemorrhagic Fevers, Viral - diagnosis Hemorrhagic Fevers, Viral - veterinary Hemorrhagic Fevers, Viral - virology Immunoglobulin G - immunology Immunoglobulin M - immunology Infectious diseases Japan Laboratories Male Maximum likelihood method Medical diagnosis Medicine and Health Sciences Methods Microbiology Phlebovirus - genetics Phlebovirus - metabolism Phylogenetics Polymerase chain reaction Research and Analysis Methods Reverse Transcriptase Polymerase Chain Reaction - methods Reverse transcription Ribonucleic acid RNA RNA viruses RNA, Viral - blood Severe Fever with Thrombocytopenia Syndrome - diagnosis Severe Fever with Thrombocytopenia Syndrome - veterinary Severe Fever with Thrombocytopenia Syndrome - virology Signs and symptoms Software Thrombocytopenia Thrombocytopenia - diagnosis Ticks Veterinary medicine Virology Viruses Wild animals Zoonoses |
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Title | Diagnostic system for the detection of severe fever with thrombocytopenia syndrome virus RNA from suspected infected animals |
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