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Rapid and Visual Detection of Coxiella burnetii Using Recombinase Polymerase Amplification Combined with Lateral Flow Strips
Coxiella burnetii, a global-distributed biological warfare agent, is the causative agent of Q fever. Correct diagnosis of Q fever is challenging and developing a fast, simple, and reliable detection method is necessary. In this study, recombinase polymerase amplification (RPA) assay combined with la...
Autores principales: | , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Hindawi
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5925019/ https://www.ncbi.nlm.nih.gov/pubmed/29850545 http://dx.doi.org/10.1155/2018/6417354 |
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author | Qi, Yong Yin, Qiong Shao, Yinxiu Li, Suqin Chen, Hongxia Shen, Wanpeng Rao, Jixian Li, Jiameng Li, Xiaoling Sun, Yu Lin, Yu Deng, Yi Zeng, Wenwen Zheng, Shulong Liu, Suyun Li, Yuexi |
author_facet | Qi, Yong Yin, Qiong Shao, Yinxiu Li, Suqin Chen, Hongxia Shen, Wanpeng Rao, Jixian Li, Jiameng Li, Xiaoling Sun, Yu Lin, Yu Deng, Yi Zeng, Wenwen Zheng, Shulong Liu, Suyun Li, Yuexi |
author_sort | Qi, Yong |
collection | PubMed |
description | Coxiella burnetii, a global-distributed biological warfare agent, is the causative agent of Q fever. Correct diagnosis of Q fever is challenging and developing a fast, simple, and reliable detection method is necessary. In this study, recombinase polymerase amplification (RPA) assay combined with lateral flow (LF) test was developed targeting 23S rRNA gene of C. burnetii Xinqiao strain. Primers and probe were designed and synthesized, with one set with high amplification efficiency screened for establishment of the method. Reaction conditions were optimized. Sensitivity, specificity, and accuracy were evaluated. The established RPA-LF reaction could be completed in 30 minutes by combining RPA at 37°C with LF at room temperature, with visually judged results. The method showed good specificity without recognizing other bacteria evaluated. It detected positive plasmid and genomic DNA at levels of 10 copies/reaction and 7 copies/reaction, respectively, levels comparable to that of real-time quantitative PCR (RT-qPCR) targeting 23S rRNA gene established previously. Both RPA-LF and RT-qPCR were used to detect C. burnetii-infected mouse samples and the results were fully consistent. The method showed superior detection performance and will provide technical support against C. burnetii in resources-limited areas. |
format | Online Article Text |
id | pubmed-5925019 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Hindawi |
record_format | MEDLINE/PubMed |
spelling | pubmed-59250192018-05-30 Rapid and Visual Detection of Coxiella burnetii Using Recombinase Polymerase Amplification Combined with Lateral Flow Strips Qi, Yong Yin, Qiong Shao, Yinxiu Li, Suqin Chen, Hongxia Shen, Wanpeng Rao, Jixian Li, Jiameng Li, Xiaoling Sun, Yu Lin, Yu Deng, Yi Zeng, Wenwen Zheng, Shulong Liu, Suyun Li, Yuexi Biomed Res Int Research Article Coxiella burnetii, a global-distributed biological warfare agent, is the causative agent of Q fever. Correct diagnosis of Q fever is challenging and developing a fast, simple, and reliable detection method is necessary. In this study, recombinase polymerase amplification (RPA) assay combined with lateral flow (LF) test was developed targeting 23S rRNA gene of C. burnetii Xinqiao strain. Primers and probe were designed and synthesized, with one set with high amplification efficiency screened for establishment of the method. Reaction conditions were optimized. Sensitivity, specificity, and accuracy were evaluated. The established RPA-LF reaction could be completed in 30 minutes by combining RPA at 37°C with LF at room temperature, with visually judged results. The method showed good specificity without recognizing other bacteria evaluated. It detected positive plasmid and genomic DNA at levels of 10 copies/reaction and 7 copies/reaction, respectively, levels comparable to that of real-time quantitative PCR (RT-qPCR) targeting 23S rRNA gene established previously. Both RPA-LF and RT-qPCR were used to detect C. burnetii-infected mouse samples and the results were fully consistent. The method showed superior detection performance and will provide technical support against C. burnetii in resources-limited areas. Hindawi 2018-04-12 /pmc/articles/PMC5925019/ /pubmed/29850545 http://dx.doi.org/10.1155/2018/6417354 Text en Copyright © 2018 Yong Qi et al. https://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Qi, Yong Yin, Qiong Shao, Yinxiu Li, Suqin Chen, Hongxia Shen, Wanpeng Rao, Jixian Li, Jiameng Li, Xiaoling Sun, Yu Lin, Yu Deng, Yi Zeng, Wenwen Zheng, Shulong Liu, Suyun Li, Yuexi Rapid and Visual Detection of Coxiella burnetii Using Recombinase Polymerase Amplification Combined with Lateral Flow Strips |
title | Rapid and Visual Detection of Coxiella burnetii Using Recombinase Polymerase Amplification Combined with Lateral Flow Strips |
title_full | Rapid and Visual Detection of Coxiella burnetii Using Recombinase Polymerase Amplification Combined with Lateral Flow Strips |
title_fullStr | Rapid and Visual Detection of Coxiella burnetii Using Recombinase Polymerase Amplification Combined with Lateral Flow Strips |
title_full_unstemmed | Rapid and Visual Detection of Coxiella burnetii Using Recombinase Polymerase Amplification Combined with Lateral Flow Strips |
title_short | Rapid and Visual Detection of Coxiella burnetii Using Recombinase Polymerase Amplification Combined with Lateral Flow Strips |
title_sort | rapid and visual detection of coxiella burnetii using recombinase polymerase amplification combined with lateral flow strips |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5925019/ https://www.ncbi.nlm.nih.gov/pubmed/29850545 http://dx.doi.org/10.1155/2018/6417354 |
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