A novel one-pot RPA-CRISPR/Cas13a diagnostic system for rapid and specific detection of HAdV infections

IF 8 1区化学 Q1 CHEMISTRY, ANALYTICAL

Sensors and Actuators B: Chemical Pub Date : 2025-03-17 DOI:10.1016/j.snb.2025.137611

Yuanyuan Li , Pan Liu , Hongyu Zhao , Lihua Liang , Minchai Chen , Xiaohan Yang

{"title":"A novel one-pot RPA-CRISPR/Cas13a diagnostic system for rapid and specific detection of HAdV infections","authors":"Yuanyuan Li , Pan Liu , Hongyu Zhao , Lihua Liang , Minchai Chen , Xiaohan Yang","doi":"10.1016/j.snb.2025.137611","DOIUrl":null,"url":null,"abstract":"<div><div>Human adenovirus (HAdV) infections are prevalent and can lead to severe respiratory, ocular, and gastrointestinal diseases, thereby posing a significant public health risk, particularly among children and immunocompromised individuals. The genetic diversity of HAdV complicates rapid and accurate detection, highlighting the necessity for efficient and straightforward diagnostic methods. This research introduces a convenient and rapid one-pot RPA-CRISPR/Cas13a diagnostic system that is both sensitive and specific for the identification of HAdV DNA. The system incorporates two detection modalities: RPA-CRISPR/Cas13a-Flu for fluorescence-based detection and RPA-CRISPR/Cas13a-LFS for visual detection. The sensitivity of the RPA-CRISPR/Cas13a-Flu method was measured at 5 copies per reaction, while the RPA-CRISPR/Cas13a-LFS demonstrated a sensitivity of 50 copies per reaction, with both methods achieving 100 % specificity. Validation against 175 clinical samples indicated that the RPA-CRISPR/Cas13a-Flu had a sensitivity of 95.95 % (95 % CI 88.75–98.61), specificity of 100 % (96.34–100), accuracy of 98.29 % (95.08–99.42), and a Kappa coefficient of 0.965 when compared to traditional qPCR methods. The RPA-CRISPR/Cas13a-LFS exhibited 81.08 % (70.71–88.38) sensitivity, 100 % (96.34–100) specificity, 92.00 % (87.02–95.17) accuracy, and a Kappa coefficient of 0.832. Our developed one-pot RPA-CRISPR/Cas13a system offers a rapid, user-friendly, and visually interpretable alternative to PCR for on-site HAdV detection, with potential applications in early diagnosis, subtyping, drug resistance detection, and portable monitoring of various infectious pathogens.</div></div>","PeriodicalId":425,"journal":{"name":"Sensors and Actuators B: Chemical","volume":"435 ","pages":"Article 137611"},"PeriodicalIF":8.0000,"publicationDate":"2025-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Sensors and Actuators B: Chemical","FirstCategoryId":"92","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0925400525003867","RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}

引用次数: 0

Abstract

Human adenovirus (HAdV) infections are prevalent and can lead to severe respiratory, ocular, and gastrointestinal diseases, thereby posing a significant public health risk, particularly among children and immunocompromised individuals. The genetic diversity of HAdV complicates rapid and accurate detection, highlighting the necessity for efficient and straightforward diagnostic methods. This research introduces a convenient and rapid one-pot RPA-CRISPR/Cas13a diagnostic system that is both sensitive and specific for the identification of HAdV DNA. The system incorporates two detection modalities: RPA-CRISPR/Cas13a-Flu for fluorescence-based detection and RPA-CRISPR/Cas13a-LFS for visual detection. The sensitivity of the RPA-CRISPR/Cas13a-Flu method was measured at 5 copies per reaction, while the RPA-CRISPR/Cas13a-LFS demonstrated a sensitivity of 50 copies per reaction, with both methods achieving 100 % specificity. Validation against 175 clinical samples indicated that the RPA-CRISPR/Cas13a-Flu had a sensitivity of 95.95 % (95 % CI 88.75–98.61), specificity of 100 % (96.34–100), accuracy of 98.29 % (95.08–99.42), and a Kappa coefficient of 0.965 when compared to traditional qPCR methods. The RPA-CRISPR/Cas13a-LFS exhibited 81.08 % (70.71–88.38) sensitivity, 100 % (96.34–100) specificity, 92.00 % (87.02–95.17) accuracy, and a Kappa coefficient of 0.832. Our developed one-pot RPA-CRISPR/Cas13a system offers a rapid, user-friendly, and visually interpretable alternative to PCR for on-site HAdV detection, with potential applications in early diagnosis, subtyping, drug resistance detection, and portable monitoring of various infectious pathogens.

查看原文本刊更多论文

求助全文

约1分钟内获得全文求助全文

来源期刊

Sensors and Actuators B: Chemical 工程技术-电化学

CiteScore

14.60

自引率

11.90%

发文量

1776

审稿时长

3.2 months

期刊介绍： Sensors & Actuators, B: Chemical is an international journal focused on the research and development of chemical transducers. It covers chemical sensors and biosensors, chemical actuators, and analytical microsystems. The journal is interdisciplinary, aiming to publish original works showcasing substantial advancements beyond the current state of the art in these fields, with practical applicability to solving meaningful analytical problems. Review articles are accepted by invitation from an Editor of the journal.