{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T17:15:26Z","timestamp":1785258926923,"version":"3.55.0"},"reference-count":35,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,7,14]],"date-time":"2021-07-14T00:00:00Z","timestamp":1626220800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a novel coronavirus that has caused the outbreak of coronavirus disease 2019 (COVID-19) all over the world. In the absence of appropriate antiviral drugs or vaccines, developing a simple, rapid, and reliable assay for SARS-CoV-2 is necessary for the prevention and control of the COVID-19 transmission.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>A novel molecular diagnosis technique, named multiplex reverse transcription loop-mediated isothermal amplification, that has been linked to a nanoparticle-based lateral flow biosensor (mRT-LAMP-LFB) was applied to detect SARS-CoV-2 based on the SARS-CoV-2 <jats:italic>RdRp<\/jats:italic> and <jats:italic>N<\/jats:italic> genes, and the mRT-LAMP products were analyzed using nanoparticle-based lateral flow biosensor. The mRT-LAMP-LFB amplification conditions, including the target RNA concentration, amplification temperature, and time were optimized. The sensitivity and specificity of the mRT-LAMP-LFB method were tested in the current study, and the mRT-LAMP-LFB assay was applied to detect the SARS-CoV-2 virus from clinical samples and artificial sputum samples.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The SARS-CoV-2 specific primers based on the <jats:italic>RdRp<\/jats:italic> and <jats:italic>N<\/jats:italic> genes were valid for the establishment of mRT-LAMP-LFB assay to detect the SARS-CoV-2 virus. The multiple-RT-LAMP amplification condition was optimized at 63\u00b0C for 30\u00a0min. The full process, including reaction preparation, viral RNA extraction, RT-LAMP, and product identification, could be achieved in 80\u00a0min. The limit of detection (LoD) of the mRT-LAMP-LFB technology was 20 copies per reaction. The specificity of mRT-LAMP-LFB detection was 100%, and no cross-reactions to other respiratory pathogens were observed.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>The mRT-LAMP-LFB technique developed in the current study is a simple, rapid, and reliable method with great specificity and sensitivity when it comes to identifying SARS-CoV-2 virus for prevention and control of the COVID-19 disease, especially in resource-constrained regions of the world.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fcimb.2021.581239","type":"journal-article","created":{"date-parts":[[2021,7,14]],"date-time":"2021-07-14T08:19:40Z","timestamp":1626250780000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":34,"title":["Rapid and Visual Detection of SARS-CoV-2 Using Multiplex Reverse Transcription Loop-Mediated Isothermal Amplification Linked With Gold Nanoparticle-Based Lateral Flow Biosensor"],"prefix":"10.3389","volume":"11","author":[{"given":"Xu","family":"Chen","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qingxue","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shijun","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Yan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bingcheng","family":"Chang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuexia","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shilei","family":"Dong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,7,14]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"e37","DOI":"10.1016\/S0140-6736(20)30309-3","article-title":"2019-Ncov Epidemic: Address Mental Health Care to Empower Society","volume":"395","author":"Bao","year":"2020","journal-title":"Lancet"},{"key":"B2","doi-asserted-by":"publisher","first-page":"69","DOI":"10.1016\/j.micinf.2020.01.004","article-title":"Pathogenicity and Transmissibility of 2019-Ncov-A Quick Overview and Comparison With Other Emerging Viruses","volume":"22","author":"Chen","year":"2020","journal-title":"Microbes Infect."},{"key":"B3","doi-asserted-by":"publisher","first-page":"30","DOI":"10.1186\/s13568-019-0754-0","article-title":"Visual and Rapid Detection of Acinetobacter Baumannii by a Multiple Cross Displacement Amplification Combined With Nanoparticles-Based Biosensor Assay","volume":"9","author":"Cheng","year":"2019","journal-title":"AMB Express."},{"key":"B4","doi-asserted-by":"publisher","first-page":"313","DOI":"10.1080\/22221751.2020.1725399","article-title":"RNA Based mNGS Approach Identifies a Novel Human Coronavirus From Two Individual Pneumonia Cases in 2019 Wuhan Outbreak","volume":"9","author":"Chen","year":"2020","journal-title":"Emerg. 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