Huifeng Du , Simone Bruno , Kalon J. Overholt , Sebastian Palacios , Hsin-Ho Huang , Carlos Barajas , Ben Gross , Cindy Lee , Haley K. Evile , Nuno Rufino de Sousa , Antonio Gigliotti Rothfuchs , Domitilla Del Vecchio
{"title":"通过基于电泳的 RNA 富集技术灵敏检测空气传播病毒的实用工艺","authors":"Huifeng Du , Simone Bruno , Kalon J. Overholt , Sebastian Palacios , Hsin-Ho Huang , Carlos Barajas , Ben Gross , Cindy Lee , Haley K. Evile , Nuno Rufino de Sousa , Antonio Gigliotti Rothfuchs , Domitilla Del Vecchio","doi":"10.1016/j.biosx.2024.100488","DOIUrl":null,"url":null,"abstract":"<div><p>Sensitive on-site virus detection is a requirement for timely action against the spread of airborne infection since ultra-low in-air viral concentrations can readily infect individuals when inhaled. Here, we consider a fieldable biosensing process that incorporates a fast RNA enrichment step in order to concentrate viral RNA in a small volume prior to RT-qPCR. The enrichment approach uses electrophoresis in an RT-qPCR-compatible buffer, and allows for concentration of RNA by nearly 5-fold within only 10 min. In order to place this performance into context, we analyzed the minimum detectable concentration of a low-cost, fieldable, biosensing process that uses electrostatic precipitation for air sampling, heating for viral RNA extraction, and then RNA enrichment, followed by RT-qPCR. With enrichment, we estimated an in-air concentration of 5,654 genome copies (gc)/m<span><math><msup><mrow></mrow><mrow><mn>3</mn></mrow></msup></math></span> with a 100% detection rate and an in-air concentration of 4,221 gc/m<span><math><msup><mrow></mrow><mrow><mn>3</mn></mrow></msup></math></span> with a 78.6% detection rate. Given that the concentrations of common viruses, such as influenza and SARS-CoV-2, in several indoor spaces are between 5,800 and 37,000 gc/m<span><math><msup><mrow></mrow><mrow><mn>3</mn></mrow></msup></math></span>, we conclude that enrichment allows a detection that is sufficiently sensitive for practical applications.</p></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"19 ","pages":"Article 100488"},"PeriodicalIF":10.6100,"publicationDate":"2024-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2590137024000529/pdfft?md5=e73dcc5dad20d173c46ff6ed390daafc&pid=1-s2.0-S2590137024000529-main.pdf","citationCount":"0","resultStr":"{\"title\":\"A fieldable process for sensitive detection of airborne viruses via electrophoresis-based RNA enrichment\",\"authors\":\"Huifeng Du , Simone Bruno , Kalon J. Overholt , Sebastian Palacios , Hsin-Ho Huang , Carlos Barajas , Ben Gross , Cindy Lee , Haley K. Evile , Nuno Rufino de Sousa , Antonio Gigliotti Rothfuchs , Domitilla Del Vecchio\",\"doi\":\"10.1016/j.biosx.2024.100488\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Sensitive on-site virus detection is a requirement for timely action against the spread of airborne infection since ultra-low in-air viral concentrations can readily infect individuals when inhaled. Here, we consider a fieldable biosensing process that incorporates a fast RNA enrichment step in order to concentrate viral RNA in a small volume prior to RT-qPCR. The enrichment approach uses electrophoresis in an RT-qPCR-compatible buffer, and allows for concentration of RNA by nearly 5-fold within only 10 min. In order to place this performance into context, we analyzed the minimum detectable concentration of a low-cost, fieldable, biosensing process that uses electrostatic precipitation for air sampling, heating for viral RNA extraction, and then RNA enrichment, followed by RT-qPCR. With enrichment, we estimated an in-air concentration of 5,654 genome copies (gc)/m<span><math><msup><mrow></mrow><mrow><mn>3</mn></mrow></msup></math></span> with a 100% detection rate and an in-air concentration of 4,221 gc/m<span><math><msup><mrow></mrow><mrow><mn>3</mn></mrow></msup></math></span> with a 78.6% detection rate. Given that the concentrations of common viruses, such as influenza and SARS-CoV-2, in several indoor spaces are between 5,800 and 37,000 gc/m<span><math><msup><mrow></mrow><mrow><mn>3</mn></mrow></msup></math></span>, we conclude that enrichment allows a detection that is sufficiently sensitive for practical applications.</p></div>\",\"PeriodicalId\":260,\"journal\":{\"name\":\"Biosensors and Bioelectronics: X\",\"volume\":\"19 \",\"pages\":\"Article 100488\"},\"PeriodicalIF\":10.6100,\"publicationDate\":\"2024-05-10\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S2590137024000529/pdfft?md5=e73dcc5dad20d173c46ff6ed390daafc&pid=1-s2.0-S2590137024000529-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biosensors and Bioelectronics: X\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2590137024000529\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"Biochemistry, Genetics and Molecular Biology\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biosensors and Bioelectronics: X","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2590137024000529","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
A fieldable process for sensitive detection of airborne viruses via electrophoresis-based RNA enrichment
Sensitive on-site virus detection is a requirement for timely action against the spread of airborne infection since ultra-low in-air viral concentrations can readily infect individuals when inhaled. Here, we consider a fieldable biosensing process that incorporates a fast RNA enrichment step in order to concentrate viral RNA in a small volume prior to RT-qPCR. The enrichment approach uses electrophoresis in an RT-qPCR-compatible buffer, and allows for concentration of RNA by nearly 5-fold within only 10 min. In order to place this performance into context, we analyzed the minimum detectable concentration of a low-cost, fieldable, biosensing process that uses electrostatic precipitation for air sampling, heating for viral RNA extraction, and then RNA enrichment, followed by RT-qPCR. With enrichment, we estimated an in-air concentration of 5,654 genome copies (gc)/m with a 100% detection rate and an in-air concentration of 4,221 gc/m with a 78.6% detection rate. Given that the concentrations of common viruses, such as influenza and SARS-CoV-2, in several indoor spaces are between 5,800 and 37,000 gc/m, we conclude that enrichment allows a detection that is sufficiently sensitive for practical applications.
期刊介绍:
Biosensors and Bioelectronics: X, an open-access companion journal of Biosensors and Bioelectronics, boasts a 2020 Impact Factor of 10.61 (Journal Citation Reports, Clarivate Analytics 2021). Offering authors the opportunity to share their innovative work freely and globally, Biosensors and Bioelectronics: X aims to be a timely and permanent source of information. The journal publishes original research papers, review articles, communications, editorial highlights, perspectives, opinions, and commentaries at the intersection of technological advancements and high-impact applications. Manuscripts submitted to Biosensors and Bioelectronics: X are assessed based on originality and innovation in technology development or applications, aligning with the journal's goal to cater to a broad audience interested in this dynamic field.