LAB ON A CHIP
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2024
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IF 6.1
Weihong Yin,Kai Hu,Bingwen Yu,Tao Zhang,Haohua Mei,Bowen Zhang,Zheyu Zou,Liping Xia,Yehong Gui,Juxin Yin,Wei Jin,Ying Mu
Escherichia coli O157:H7 is a major foodborne pathogen that poses a significant threat to food safety and human health. Rapid and sensitive detection of viable Escherichia coli O157:H7 can effectively prevent food poisoning. Here, we developed a microwell-confined and propidium monoazide-assisted digital CRISPR microfluidic platform for rapid and sensitive detection of viable Escherichia coli O157:H7 in food samples. This reaction time significantly reduced by minimizing the microwell volume, yielding qualitative results in 5 min and absolute quantitative results in 15 min. With the assistance of propidium monoazide, this platform can eliminate the interference from 99% of dead Escherichia coli O157:H7. The direct lysis method obviates the need for the complex nucleic acid extraction process, offering a limit of detection of 3.6×101 CFU/mL within 30 min. Our results demonstrated that the platform provides a powerful tool for rapid detection of Escherichia coli O157:H7 and provides reliable guidance for food safety testing.
研究领域:
微生物学
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分子生物学
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食品科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2024
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IF 6.7
Weihong Yin,Kai Hu,Yunxing Yang,Jianjian Zhuang,Zheyu Zou,Yuanjie Suo,Liping Xia,Jiale Li,Yehong Gui,Haohua Mei,Juxin Yin,Tao Zhang,Ying Mu
Escherichia coli O157:H7 (E. coli O157:H7) is a prominent pathogenic bacterium that poses serious risks to food safety and public health. Rapid and accurate detection of live E. coli O157:H7 is of great importance in food quality monitoring and clinical diagnosis. Here, we report a propidium monoazide-assisted nonamplification digital CRISPR/Cas12a assay for sensitive and rapid detection of live E. coli O157:H7. The incorporation of propidium monoazide into the method enables the selective detection of live bacteria by eliminating 98% of interference from the dead bacterial nucleic acid. Implemented on microfluidic digital chips, this method can achieve absolute quantification of nonamplified nucleic acid. The entire detection process of live bacteria can be completed within 120 min without the need for establishing a standard curve, and the sensitivity of the method reaches 1.2 × 103 CFU/mL. The method was validated using various samples, yielding results consistent with the plate counting method (Pearson’s r = 0.9490). Consequently, this method holds significant potential for applications in fields requiring live bacterial detection.
研究领域:
生物技术
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分子生物学
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食品科学
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公共卫生
✓ 已验证
产品关联
DOI
PubMed
高影响因子
LWT-FOOD SCIENCE AND TECHNOLOGY
·
2023
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IF 6
Junyan Liu,Tengyi Huang,Riyi Xu,Zhufang Xiang,Thanapop Soteyome,Xiangyi Chen,Qianfei Zhang,Qiurong Huang,Zixu Wu,Yunyi Huang,Gongliang Liu,Chunyun Qu,Gamini Seneviratne,Birthe V. Kjellerup,Zhenbo Xu
Escherichia coli O157:H7 is a major foodborne pathogen causing food poisoning. Standard detection method for foodborne E. coli is plate counting, which is time consuming and omits VBNC cells. PSR is an isothermal amplification technique identifying trace amount DNA. However, its effectiveness is influenced by residual DNA from dead cells. This study aimed at designing a PMA-PSR system for accurate determination of viable cells of E. coli O157:H7. Firstly, PSR was optimized in fluorescence indicator, betaine concentration and reaction time. The optimized system could successfully identify rfbE, stx1 and stx2 genes within 45 min at 65 °C. Secondly, sensitivity and specificity were tested in pure culture and different food samples. With 100% specificity, PSR could determine DNA amount as low as 1.12 pg/μL. Thirdly, a PMA-PSR system was designed for accurate determination of viable cells. Cells were treated with 5 μg/mL PMA to remove residual DNA from dead cells. Pretreated cells were adapted to PSR, thus viable cells were specifically detected. Fourthly, the system was applied in food samples with VBNC cells. Viable cells could be accurately determined in accordance with regular assay. In conclusion, the PMA-PSR system was an efficient tool for accurate determination of E. coli O157:H7 viable cells.
研究领域:
微生物学
·
食品科学
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）