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Development of an RT-LAMP Assay for the Rapid Detection of SFTS Virus
Detection of severe fever with thrombocytopenia syndrome (SFTS) virus (SFTSV) during the early phase of the disease is important for appropriate treatment, infection control, and prevention of further transmission. The reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a nucle...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8073756/ https://www.ncbi.nlm.nih.gov/pubmed/33923720 http://dx.doi.org/10.3390/v13040693 |
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author | Sano, Shiori Fukushi, Shuetsu Yamada, Souichi Harada, Shizuko Kinoshita, Hitomi Sugimoto, Satoko Yoshikawa, Tomoki Kurosu, Takeshi Takamatsu, Yuki Shimojima, Masayuki Toda, Shoichi Hamada, Yuka Fujisawa, Naoki Sugimoto, Takayuki Saijo, Masayuki |
author_facet | Sano, Shiori Fukushi, Shuetsu Yamada, Souichi Harada, Shizuko Kinoshita, Hitomi Sugimoto, Satoko Yoshikawa, Tomoki Kurosu, Takeshi Takamatsu, Yuki Shimojima, Masayuki Toda, Shoichi Hamada, Yuka Fujisawa, Naoki Sugimoto, Takayuki Saijo, Masayuki |
author_sort | Sano, Shiori |
collection | PubMed |
description | Detection of severe fever with thrombocytopenia syndrome (SFTS) virus (SFTSV) during the early phase of the disease is important for appropriate treatment, infection control, and prevention of further transmission. The reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a nucleic acid amplification method that amplifies the target sequence under isothermal conditions. Here, we developed an RT-LAMP with a novel primer/probe set targeting a conserved region of the SFTSV L segment after extraction of viral RNA (standard RT-LAMP). Both the Chinese and Japanese SFTSV strains, including various genotypes, were detected by the standard RT-LAMP. We also performed RT-LAMP using the same primer/probe set but without the viral RNA extraction step (called simplified RT-LAMP) and evaluated the diagnostic efficacy. The sensitivity and specificity of the simplified RT-LAMP were 84.9% (45/53) and 89.5% (2/19), respectively. The simplified RT-LAMP can detect SFTSV in human sera containing >10(3.5) copies/mL viral RNA. The two RT-LAMP positive but quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) negative samples were positive in the conventional RT-PCR, suggesting that there was no false positive reaction in the RT-LAMP. Both the standard and simplified RT-LAMP are useful for detecting the SFTSV genome in patients during the early phase of the disease. |
format | Online Article Text |
id | pubmed-8073756 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-80737562021-04-27 Development of an RT-LAMP Assay for the Rapid Detection of SFTS Virus Sano, Shiori Fukushi, Shuetsu Yamada, Souichi Harada, Shizuko Kinoshita, Hitomi Sugimoto, Satoko Yoshikawa, Tomoki Kurosu, Takeshi Takamatsu, Yuki Shimojima, Masayuki Toda, Shoichi Hamada, Yuka Fujisawa, Naoki Sugimoto, Takayuki Saijo, Masayuki Viruses Article Detection of severe fever with thrombocytopenia syndrome (SFTS) virus (SFTSV) during the early phase of the disease is important for appropriate treatment, infection control, and prevention of further transmission. The reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a nucleic acid amplification method that amplifies the target sequence under isothermal conditions. Here, we developed an RT-LAMP with a novel primer/probe set targeting a conserved region of the SFTSV L segment after extraction of viral RNA (standard RT-LAMP). Both the Chinese and Japanese SFTSV strains, including various genotypes, were detected by the standard RT-LAMP. We also performed RT-LAMP using the same primer/probe set but without the viral RNA extraction step (called simplified RT-LAMP) and evaluated the diagnostic efficacy. The sensitivity and specificity of the simplified RT-LAMP were 84.9% (45/53) and 89.5% (2/19), respectively. The simplified RT-LAMP can detect SFTSV in human sera containing >10(3.5) copies/mL viral RNA. The two RT-LAMP positive but quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) negative samples were positive in the conventional RT-PCR, suggesting that there was no false positive reaction in the RT-LAMP. Both the standard and simplified RT-LAMP are useful for detecting the SFTSV genome in patients during the early phase of the disease. MDPI 2021-04-16 /pmc/articles/PMC8073756/ /pubmed/33923720 http://dx.doi.org/10.3390/v13040693 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Sano, Shiori Fukushi, Shuetsu Yamada, Souichi Harada, Shizuko Kinoshita, Hitomi Sugimoto, Satoko Yoshikawa, Tomoki Kurosu, Takeshi Takamatsu, Yuki Shimojima, Masayuki Toda, Shoichi Hamada, Yuka Fujisawa, Naoki Sugimoto, Takayuki Saijo, Masayuki Development of an RT-LAMP Assay for the Rapid Detection of SFTS Virus |
title | Development of an RT-LAMP Assay for the Rapid Detection of SFTS Virus |
title_full | Development of an RT-LAMP Assay for the Rapid Detection of SFTS Virus |
title_fullStr | Development of an RT-LAMP Assay for the Rapid Detection of SFTS Virus |
title_full_unstemmed | Development of an RT-LAMP Assay for the Rapid Detection of SFTS Virus |
title_short | Development of an RT-LAMP Assay for the Rapid Detection of SFTS Virus |
title_sort | development of an rt-lamp assay for the rapid detection of sfts virus |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8073756/ https://www.ncbi.nlm.nih.gov/pubmed/33923720 http://dx.doi.org/10.3390/v13040693 |
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