FOOD CONTROL
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2025
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IF 6.3
Yixiang Sun,Liren Zhang,Huimin Wu,Hongjun Chen,Huijie Hu,Chongyu Zhang,Xiaoqiang Li,Yuan Li,Yimai Wang,Liqiang Luo,Yizhi Song
Methicillin-resistant Staphylococcus aureus (MRSA) has emerged as a significant pathogen of global concern, presenting substantial public health risks through foodborne transmission and contributing to elevated morbidity and mortality rates worldwide. The critical need for precise and timely identification of MRSA in food matrices has become increasingly paramount for effective public health protection. While numerous CRISPR-based detection platforms for MRSA have been recently developed, their widespread implementation has been hindered by intricate multi-step protocols and dependency on sophisticated instrumentation. In this study, we developed a direct, accurate and visual detection method for MRSA — the RPA-Cas12a-LFS method. This method comprises two main components: (1) the direct one-pot RPA-Cas12a system, which integrates bacterial lysis, RPA nucleic acid amplification, and CRISPR-Cas12a nucleic acid detection into a single step performed simultaneously at a constant temperature, and (2) the streptavidin-gold nanoparticles (SA-AuNP)-based CRISPR-specific lateral flow strip (LFS). By eliminating the need for nucleic acid extraction, this method significantly simplifies the experimental procedure and reduces the risk of cross-contamination. Through systematic optimization, this method demonstrated exceptional performance, enabling direct and specific identification of MRSA at remarkably low concentrations (10 CFU/mL) within 60 min in various milk samples. This advanced detection method, characterized by its direct sample processing, exceptional accuracy, visual interpretability, cost-efficiency, and minimal equipment requirements, is particularly suitable for on-site and real-time monitoring of pathogenic bacteria in the food industry.
研究领域:
微生物学
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食品安全
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分子诊断
✓ 已验证
产品关联
DOI
高影响因子
ANALYTICA CHIMICA ACTA
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2023
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IF 6.2
Ling Deng,Xiaoyi He,Ke Liu,Yuanli Li,Han Xia,Hang Qian,Xiaoxue Lu,Xuhu Mao,Yang Xiang
Burkholderia pseudomallei is the causative agent of melioidosis, a potentially life-threatening infectious disease, and poses public health risks in endemic areas. Due to the high mortality, intrinsic antibiotic resistance , and atypical manifestations, establishing a rapid, accurate, and sensitive identification of B. pseudomallei enables earlier diagnosis, proper treatments, and better outcomes of melioidosis. Herein, we present a O ne- P ot C RISPR-integrated assay for I nstant and V isual D etection (termed OPC-IVD) of B. pseudomallei . The integration of recombinase polymerase amplification and CRISPR-Cas12a recognition-activated trans -cleavage, achieved a true all-in-one single-tube reaction system, initiating the amplification and cleavage simultaneously, which realized a facile sample-to-answer assay. This approach could be performed with simplified DNA extraction and completed around 30 min by holding the reaction tube in the hand. The detection limit of our OPC-IVD was determined to be 2.19 copy/uL of plasmid DNA, 12.5 CFU/mL of B. pseudomallei , and 61.5 CFU/mL of bacteria in spiked blood samples, respectively. Furthermore, the introduction of internal amplification control effectively reduced the occurrence of false negatives, which was incorporated in the reaction system, and amplified simultaneously with the target and read by naked eyes. The assay exhibited 100% accuracy when evaluated in clinical isolates and samples. The streamlined workflow of our OPC-IVD of B. pseudomallei enables a field-deployable, instrument-free, and ultra-fast approach that can be utilized by non-expert personnel in the field of molecular diagnosis of melioidosis especially in under-resourced setting.
研究领域:
医学诊断
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分子生物学
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传染病学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Small Methods
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2025
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IF 9.1
Xiaoying Han,Juxin Yin,Yu Wang,Jianjian Zhuang,Kai Hu,Yehong Gui,Haohua Mei,Jizhi Tong,Ying Mu
The rapid, precise, and automated diagnosis of infectious diseases is crucial for effective disease management and control. Herein, the integrated portable and automatic digital detection system (IPADS), a novel diagnostic platform for nucleic acid detection is introduced. The device employs the hybrid magnetic system (HMS), which uses an electromagnet and a movable permanent magnet to modulate the magnetic field and control bead movement, increasing nucleic acid extraction efficiency to over 80%, while simplifying the traditional labor-intensive process and enabling quick, low-risk sample processing. Additionally, a disposable cartridge is designed for integrated HMS based preprocessing, with detection performed using digital RPA-Cas12a, enabling rapid, enclosed, and automation-friendly detection across a dynamic range spanning five orders of magnitude, with a sensitivity as low as 100 copies mL −1 in serum samples. An automated platform further optimizes workflow. As a proof of concept, IPADS is applied to detect hepatitis B virus (HBV) DNA in 20 clinical serum samples, demonstrating high concordance with gold-standard quantitative PCR (qPCR) methods. These results validate the potential of IPADS as a reliable point-of-care testing solution.
研究领域:
医学诊断
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分子生物学
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生物医学工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）