Yangdao Wei , Zhenzhen Tao , Lu Wan , Chengli Zong , Jiajia Wu , Xiao Tan , Buhua Wang , Zixuan Guo , Ling Zhang , Haoyu Yuan , Peng Wang , Zhiqing Yang , Yi Wan
{"title":"基于适体体的Cas14a1生物传感器用于无扩增活致病性检测","authors":"Yangdao Wei , Zhenzhen Tao , Lu Wan , Chengli Zong , Jiajia Wu , Xiao Tan , Buhua Wang , Zixuan Guo , Ling Zhang , Haoyu Yuan , Peng Wang , Zhiqing Yang , Yi Wan","doi":"10.1016/j.bios.2022.114282","DOIUrl":null,"url":null,"abstract":"<div><p><span>CRISPR-Cas systems have been employed to detect a large variety of pathogenic microorganisms by simply changing the guide RNA<span><span> sequence. However, these platforms usually rely on nucleic acid extraction and amplification to achieve good sensitivity. Herein, we developed a new platform for the highly specific and </span>sensitive detection of live </span></span><span><em>staphylococcus aureus</em></span> (<em>S. aureus</em>) based on an <strong><u>A</u></strong>ptamer-based <strong><u>Cas</u></strong>14a1 <strong><u>B</u></strong>iosensor (<strong>ACasB</strong>), without the need for nucleic acid extraction or amplification. First, the <em>S. aureus</em><span> specific aptamer was hybrid with a blocker DNA. After the live </span><em>S. aureus</em> was added, the blocker can be released upon bacteria-aptamer binding. Finally, the released blocker can activate Cas14a1 protein by binding with the sgRNA to generate a change of fluorescent intensity. The ACasB indicates high specificity and sensitivity: it can directly distinguish 400 CFU/ml live <em>S. aureus</em> cells. Comparable to qPCR, the Cas14a1-aptamer biosensor can detect <em>S. aureus</em> with 100% accuracy in complex samples. Therefore, this ACasB for the on-site detection of live <em>S. aureus</em> can broaden its applications in food safety and environmental monitoring.</p></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":null,"pages":null},"PeriodicalIF":10.7000,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"27","resultStr":"{\"title\":\"Aptamer-based Cas14a1 biosensor for amplification-free live pathogenic detection\",\"authors\":\"Yangdao Wei , Zhenzhen Tao , Lu Wan , Chengli Zong , Jiajia Wu , Xiao Tan , Buhua Wang , Zixuan Guo , Ling Zhang , Haoyu Yuan , Peng Wang , Zhiqing Yang , Yi Wan\",\"doi\":\"10.1016/j.bios.2022.114282\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p><span>CRISPR-Cas systems have been employed to detect a large variety of pathogenic microorganisms by simply changing the guide RNA<span><span> sequence. However, these platforms usually rely on nucleic acid extraction and amplification to achieve good sensitivity. Herein, we developed a new platform for the highly specific and </span>sensitive detection of live </span></span><span><em>staphylococcus aureus</em></span> (<em>S. aureus</em>) based on an <strong><u>A</u></strong>ptamer-based <strong><u>Cas</u></strong>14a1 <strong><u>B</u></strong>iosensor (<strong>ACasB</strong>), without the need for nucleic acid extraction or amplification. First, the <em>S. aureus</em><span> specific aptamer was hybrid with a blocker DNA. After the live </span><em>S. aureus</em> was added, the blocker can be released upon bacteria-aptamer binding. Finally, the released blocker can activate Cas14a1 protein by binding with the sgRNA to generate a change of fluorescent intensity. The ACasB indicates high specificity and sensitivity: it can directly distinguish 400 CFU/ml live <em>S. aureus</em> cells. Comparable to qPCR, the Cas14a1-aptamer biosensor can detect <em>S. aureus</em> with 100% accuracy in complex samples. Therefore, this ACasB for the on-site detection of live <em>S. aureus</em> can broaden its applications in food safety and environmental monitoring.</p></div>\",\"PeriodicalId\":259,\"journal\":{\"name\":\"Biosensors and Bioelectronics\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":10.7000,\"publicationDate\":\"2022-09-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"27\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biosensors and Bioelectronics\",\"FirstCategoryId\":\"1\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0956566322003220\",\"RegionNum\":1,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"BIOPHYSICS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biosensors and Bioelectronics","FirstCategoryId":"1","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0956566322003220","RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOPHYSICS","Score":null,"Total":0}
Aptamer-based Cas14a1 biosensor for amplification-free live pathogenic detection
CRISPR-Cas systems have been employed to detect a large variety of pathogenic microorganisms by simply changing the guide RNA sequence. However, these platforms usually rely on nucleic acid extraction and amplification to achieve good sensitivity. Herein, we developed a new platform for the highly specific and sensitive detection of live staphylococcus aureus (S. aureus) based on an Aptamer-based Cas14a1 Biosensor (ACasB), without the need for nucleic acid extraction or amplification. First, the S. aureus specific aptamer was hybrid with a blocker DNA. After the live S. aureus was added, the blocker can be released upon bacteria-aptamer binding. Finally, the released blocker can activate Cas14a1 protein by binding with the sgRNA to generate a change of fluorescent intensity. The ACasB indicates high specificity and sensitivity: it can directly distinguish 400 CFU/ml live S. aureus cells. Comparable to qPCR, the Cas14a1-aptamer biosensor can detect S. aureus with 100% accuracy in complex samples. Therefore, this ACasB for the on-site detection of live S. aureus can broaden its applications in food safety and environmental monitoring.
期刊介绍:
Biosensors & Bioelectronics, along with its open access companion journal Biosensors & Bioelectronics: X, is the leading international publication in the field of biosensors and bioelectronics. It covers research, design, development, and application of biosensors, which are analytical devices incorporating biological materials with physicochemical transducers. These devices, including sensors, DNA chips, electronic noses, and lab-on-a-chip, produce digital signals proportional to specific analytes. Examples include immunosensors and enzyme-based biosensors, applied in various fields such as medicine, environmental monitoring, and food industry. The journal also focuses on molecular and supramolecular structures for enhancing device performance.