A ERA/Cas12f1_ge4.1 biosensor for rapid, sensitive, and cost-effective detection of Chlamydia psittaci via fluorescence and lateral flow assays

IF 6.1 1区化学 Q1 CHEMISTRY, ANALYTICAL

Talanta Pub Date : 2025-01-24 DOI:10.1016/j.talanta.2025.127615

Tingqing Jiang , Guijiang Wei , Mei Lin , Sheping Zhang , Lintao Zou , Xuan Zhou , Zhongliang Deng

{"title":"A ERA/Cas12f1_ge4.1 biosensor for rapid, sensitive, and cost-effective detection of Chlamydia psittaci via fluorescence and lateral flow assays","authors":"Tingqing Jiang , Guijiang Wei , Mei Lin , Sheping Zhang , Lintao Zou , Xuan Zhou , Zhongliang Deng","doi":"10.1016/j.talanta.2025.127615","DOIUrl":null,"url":null,"abstract":"<div><div>Rapid and accurate detection of <em>Chlamydia psittaci</em>, the causative agent of psittacosis, is crucial for both human and animal health but presents significant challenges, particularly in grassroots health institutions. Our previous PDTCTR fluorescence sensing platform, which combined the engineered Cas12f1_ge4.1 system with recombinase polymerase amplification (RPA), significantly enhanced detection efficiency. However, its requirement for specialized equipment, costly RPA reagents, and absence of visual output restricted its practical application in such environments. To address these limitations, we developed the ERA/Cas12f1_ge4.1 system, integrating Cas12f1_ge4.1 with the cost-effective Enzymatic Recombinase Amplification (ERA). This system enables sensitive detection of <em>Chlamydia psittaci</em> double-stranded DNA within 50 min through both fluorescence and colloidal gold lateral flow assay strips. The platform achieves detection limits of 10 copies/μL for fluorescence and 100 copies/μL for lateral flow. Clinical validation involving 93 parrot samples demonstrated high performance in both detection modes. Fluorescence detection achieved 95.4 % sensitivity, 100 % specificity, a 100 % positive predictive value (PPV), and a 90.3 % negative predictive value (NPV). Meanwhile, the lateral flow assay exhibited 92.3 % sensitivity, 100 % specificity, 100 % PPV, and an 84.8 % NPV. The ERA/Cas12f1_ge4.1 system offers a rapid, accurate, cost-effective, and visually interpretable diagnostic tool suitable for both laboratory and community health centers. This advancement holds significant potential for improving psittacosis diagnosis and control, particularly in resource-limited environments.</div></div>","PeriodicalId":435,"journal":{"name":"Talanta","volume":"287 ","pages":"Article 127615"},"PeriodicalIF":6.1000,"publicationDate":"2025-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Talanta","FirstCategoryId":"92","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0039914025001018","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

Rapid and accurate detection of Chlamydia psittaci, the causative agent of psittacosis, is crucial for both human and animal health but presents significant challenges, particularly in grassroots health institutions. Our previous PDTCTR fluorescence sensing platform, which combined the engineered Cas12f1_ge4.1 system with recombinase polymerase amplification (RPA), significantly enhanced detection efficiency. However, its requirement for specialized equipment, costly RPA reagents, and absence of visual output restricted its practical application in such environments. To address these limitations, we developed the ERA/Cas12f1_ge4.1 system, integrating Cas12f1_ge4.1 with the cost-effective Enzymatic Recombinase Amplification (ERA). This system enables sensitive detection of Chlamydia psittaci double-stranded DNA within 50 min through both fluorescence and colloidal gold lateral flow assay strips. The platform achieves detection limits of 10 copies/μL for fluorescence and 100 copies/μL for lateral flow. Clinical validation involving 93 parrot samples demonstrated high performance in both detection modes. Fluorescence detection achieved 95.4 % sensitivity, 100 % specificity, a 100 % positive predictive value (PPV), and a 90.3 % negative predictive value (NPV). Meanwhile, the lateral flow assay exhibited 92.3 % sensitivity, 100 % specificity, 100 % PPV, and an 84.8 % NPV. The ERA/Cas12f1_ge4.1 system offers a rapid, accurate, cost-effective, and visually interpretable diagnostic tool suitable for both laboratory and community health centers. This advancement holds significant potential for improving psittacosis diagnosis and control, particularly in resource-limited environments.

Abstract Image

查看原文本刊更多论文

ERA/Cas12f1_ge4.1生物传感器，用于快速、灵敏、经济高效地通过荧光和侧流分析检测猪瘟衣原体。

快速、准确地检测鹦鹉热病原体鹦鹉热衣原体对人类和动物健康都至关重要，但也带来了重大挑战，特别是在基层卫生机构。我们之前的PDTCTR荧光传感平台将工程Cas12f1_ge4.1系统与重组酶聚合酶扩增（RPA）相结合，显著提高了检测效率。然而，它对专用设备、昂贵的RPA试剂和缺乏视觉输出的要求限制了它在这种环境中的实际应用。为了解决这些限制，我们开发了ERA/Cas12f1_ge4.1系统，将Cas12f1_ge4.1与具有成本效益的酶重组酶扩增（ERA）相结合。该系统可在50分钟内通过荧光和胶体金横向流动试验条对鹦鹉热衣原体双链DNA进行灵敏检测。荧光检测限为10拷贝/μL，侧流检测限为100拷贝/μL。涉及93个鹦鹉样本的临床验证表明，这两种检测模式都具有很高的性能。荧光检测灵敏度为95.4%，特异性为100%，阳性预测值为100%，阴性预测值为90.3%。同时，横向流动试验的灵敏度为92.3%，特异性为100%，PPV为100%，NPV为84.8%。ERA/Cas12f1_ge4.1系统为实验室和社区卫生中心提供了一种快速、准确、具有成本效益和视觉可解释的诊断工具。这一进展对改善鹦鹉热的诊断和控制具有重大潜力，特别是在资源有限的环境中。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Talanta 化学-分析化学

CiteScore

12.30

自引率

4.90%

发文量

861

审稿时长

29 days

期刊介绍： Talanta provides a forum for the publication of original research papers, short communications, and critical reviews in all branches of pure and applied analytical chemistry. Papers are evaluated based on established guidelines, including the fundamental nature of the study, scientific novelty, substantial improvement or advantage over existing technology or methods, and demonstrated analytical applicability. Original research papers on fundamental studies, and on novel sensor and instrumentation developments, are encouraged. Novel or improved applications in areas such as clinical and biological chemistry, environmental analysis, geochemistry, materials science and engineering, and analytical platforms for omics development are welcome. Analytical performance of methods should be determined, including interference and matrix effects, and methods should be validated by comparison with a standard method, or analysis of a certified reference material. Simple spiking recoveries may not be sufficient. The developed method should especially comprise information on selectivity, sensitivity, detection limits, accuracy, and reliability. However, applying official validation or robustness studies to a routine method or technique does not necessarily constitute novelty. Proper statistical treatment of the data should be provided. Relevant literature should be cited, including related publications by the authors, and authors should discuss how their proposed methodology compares with previously reported methods.