Shuwen Qian, Dr. Erin M. McConnell, Meghan Rothenbroker, Jimmy Gu, Simina Alungulesa, Louis Godbout, Prof. Yingfu Li
{"title":"DNAzyme/珠状荧光法检测冷却塔水样中的嗜肺军团菌","authors":"Shuwen Qian, Dr. Erin M. McConnell, Meghan Rothenbroker, Jimmy Gu, Simina Alungulesa, Louis Godbout, Prof. Yingfu Li","doi":"10.1002/anse.202300020","DOIUrl":null,"url":null,"abstract":"<p><i>Legionella pneumophila</i> is the causative agent behind the deadly waterborne disease Legionnaires’, which is commonly transmitted by the spread of contaminated droplets from cooling tower water samples. The lack of effective detection methods presents a challenge for <i>L. pneumophila</i> outbreak control. Previously, an RNA-cleaving DNAzyme called LP1 was reported to specifically target <i>L. pneumophila</i>. In this study, LP1 was immobilized onto agarose beads via streptavidin-biotin interaction to develop a bead-based fluorescence assay for <i>L. pneumophila</i> detection. This bead-based assay demonstrated excellent stability and functionality in various cooling tower water samples. To improve <i>L. pneumophila</i> monitoring in real-world samples, a lysozyme treatment was used to enhance <i>L. pneumophila</i> recognition. The limit of detection of this DNAzyme-based bead assay can reach 10<sup>3</sup> CFUs in cell-spiked cooling tower water samples without cell culturing or signal amplification steps.</p>","PeriodicalId":72192,"journal":{"name":"Analysis & sensing","volume":null,"pages":null},"PeriodicalIF":3.4000,"publicationDate":"2023-04-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://chemistry-europe.onlinelibrary.wiley.com/doi/epdf/10.1002/anse.202300020","citationCount":"0","resultStr":"{\"title\":\"Detecting Legionella pneumophila in Cooling Tower Water Samples with a DNAzyme/Bead-Based Fluorescence Assay\",\"authors\":\"Shuwen Qian, Dr. Erin M. McConnell, Meghan Rothenbroker, Jimmy Gu, Simina Alungulesa, Louis Godbout, Prof. Yingfu Li\",\"doi\":\"10.1002/anse.202300020\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><i>Legionella pneumophila</i> is the causative agent behind the deadly waterborne disease Legionnaires’, which is commonly transmitted by the spread of contaminated droplets from cooling tower water samples. The lack of effective detection methods presents a challenge for <i>L. pneumophila</i> outbreak control. Previously, an RNA-cleaving DNAzyme called LP1 was reported to specifically target <i>L. pneumophila</i>. In this study, LP1 was immobilized onto agarose beads via streptavidin-biotin interaction to develop a bead-based fluorescence assay for <i>L. pneumophila</i> detection. This bead-based assay demonstrated excellent stability and functionality in various cooling tower water samples. To improve <i>L. pneumophila</i> monitoring in real-world samples, a lysozyme treatment was used to enhance <i>L. pneumophila</i> recognition. The limit of detection of this DNAzyme-based bead assay can reach 10<sup>3</sup> CFUs in cell-spiked cooling tower water samples without cell culturing or signal amplification steps.</p>\",\"PeriodicalId\":72192,\"journal\":{\"name\":\"Analysis & sensing\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":3.4000,\"publicationDate\":\"2023-04-27\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://chemistry-europe.onlinelibrary.wiley.com/doi/epdf/10.1002/anse.202300020\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Analysis & sensing\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/anse.202300020\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"CHEMISTRY, ANALYTICAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Analysis & sensing","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/anse.202300020","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}
Detecting Legionella pneumophila in Cooling Tower Water Samples with a DNAzyme/Bead-Based Fluorescence Assay
Legionella pneumophila is the causative agent behind the deadly waterborne disease Legionnaires’, which is commonly transmitted by the spread of contaminated droplets from cooling tower water samples. The lack of effective detection methods presents a challenge for L. pneumophila outbreak control. Previously, an RNA-cleaving DNAzyme called LP1 was reported to specifically target L. pneumophila. In this study, LP1 was immobilized onto agarose beads via streptavidin-biotin interaction to develop a bead-based fluorescence assay for L. pneumophila detection. This bead-based assay demonstrated excellent stability and functionality in various cooling tower water samples. To improve L. pneumophila monitoring in real-world samples, a lysozyme treatment was used to enhance L. pneumophila recognition. The limit of detection of this DNAzyme-based bead assay can reach 103 CFUs in cell-spiked cooling tower water samples without cell culturing or signal amplification steps.