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<p>Infections caused by <i>Escherichia coli</i> (<i>E. coli</i>) O157:H7 represent a significant global public health challenge. Consequently, the early detection of <i>E. coli</i> O157:H7 is critical for ensuring food safety and safeguarding human health. This study developed real-time fluorescent and on-site visual saltatory rolling circle amplification (SRCA) assays to enable closed-tube, rapid, and highly sensitive detection of <i>E. coli</i> O157:H7, thereby substantially reducing the risk of false-positive results. Positive and negative samples were effectively distinguished through real-time fluorescent curve analysis or direct visual assessment by naked eye. The assays demonstrated a detection sensitivity of 3.2&#xa0;fg/μL for <i>E. coli</i> O157:H7 genomic DNA, representing a 100-fold improvement over conventional real-time quantitative PCR (qPCR). Furthermore, the limit of detection in spiked milk samples was 6.7&#xa0;CFU/mL, which was 100 times lower than that achieved by qPCR. The relative sensitivity, specificity and accuracy of the two methods were 100%, 98.28% and 98.33%, respectively. In summary, the SRCA-based real-time fluorescent and visual detection strategies developed in this study exhibit high sensitivity and specificity. Their advantages including simplicity, rapidity, and cost-effectiveness make them suitable for diverse detection scenarios. These assays thus provide a novel and efficient technical solution for the rapid identification of <i>E. coli</i> O157:H7, with significant implications for food safety and public health surveillance.</p>

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Closed-tube rapid and sensitive detection of Escherichia coli O157:H7 by real-time fluorescent or on-site visual SRCA in food

  • 期刊论文

  • 发表日期：

    2026年1月21日

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  • Shuai Lei,
  • Siya Yin,
  • Guibin Cui,
  • Lina Pang,
  • Guiping Sun,
  • Wei Zhang,
  • Qian Yang

摘要

Infections caused by Escherichia coli (E. coli) O157:H7 represent a significant global public health challenge. Consequently, the early detection of E. coli O157:H7 is critical for ensuring food safety and safeguarding human health. This study developed real-time fluorescent and on-site visual saltatory rolling circle amplification (SRCA) assays to enable closed-tube, rapid, and highly sensitive detection of E. coli O157:H7, thereby substantially reducing the risk of false-positive results. Positive and negative samples were effectively distinguished through real-time fluorescent curve analysis or direct visual assessment by naked eye. The assays demonstrated a detection sensitivity of 3.2 fg/μL for E. coli O157:H7 genomic DNA, representing a 100-fold improvement over conventional real-time quantitative PCR (qPCR). Furthermore, the limit of detection in spiked milk samples was 6.7 CFU/mL, which was 100 times lower than that achieved by qPCR. The relative sensitivity, specificity and accuracy of the two methods were 100%, 98.28% and 98.33%, respectively. In summary, the SRCA-based real-time fluorescent and visual detection strategies developed in this study exhibit high sensitivity and specificity. Their advantages including simplicity, rapidity, and cost-effectiveness make them suitable for diverse detection scenarios. These assays thus provide a novel and efficient technical solution for the rapid identification of E. coli O157:H7, with significant implications for food safety and public health surveillance.