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Development of the Saltatory Rolling Circle Amplification Assay for Rapid and Visual Detection of Alicyclobacillus acidoterrestris in Apple Juice
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2020-04-06 , DOI: 10.1021/acs.jafc.0c00061 Ning Yuan 1 , Yunzhe Zhang 2 , Hui Xu 1 , Zhijun Zhou 3 , Xin Lu 1 , Tingting Chen 2 , Qian Yang 1 , Jianxin Tan 2 , Wei Zhang 1, 2, 4
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2020-04-06 , DOI: 10.1021/acs.jafc.0c00061 Ning Yuan 1 , Yunzhe Zhang 2 , Hui Xu 1 , Zhijun Zhou 3 , Xin Lu 1 , Tingting Chen 2 , Qian Yang 1 , Jianxin Tan 2 , Wei Zhang 1, 2, 4
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A novel nucleic acid isothermal amplification method based on saltatory rolling circle amplification (SRCA) for rapid and visual detection of Alicyclobacillus acidoterrestris in apple juice was established. Fourteen A. acidoterrestris strains and 44 non-A. acidoterrestris strains were used to confirm the specificity. The sensitivity of SRCA was 4.5 × 101 CFU/mL by observing the white precipitate with the naked eye, while it was 4.5 × 100 CFU/mL by fluorescence visualization. The detection limit of SRCA in artificially inoculated apple juice was 7.1 × 101 and 7.1 × 100 CFU/mL via visualization of the white precipitate and fluorescence, respectively. Compared with the traditional PCR method, SRCA exhibited at least a 100-fold higher sensitivity and 100-fold lower detection limit. Seventy samples were investigated for A. acidoterrestris contamination, and the results showed 100% sensitivity, 97.01% specificity, and 97.14% accuracy compared with those by the conventional microbiological cultivation method. Overall, this method is a potentially useful tool for visual and rapid detection of A. acidoterrestris.
中文翻译：
快速和目视检测苹果汁中酸脂环酸杆菌的咸化滚动圆扩增法的发展
建立了一种基于盐化滚环扩增（SRCA）的核酸等温扩增新方法，用于苹果汁中酸热脂环酸杆菌的快速直观检测。使用十四个酸曲霉菌株和44个非酸曲霉菌株来确认特异性。通过肉眼观察白色沉淀物，SRCA的灵敏度为4.5×10 1 CFU / mL，而通过荧光可视化为4.5×10 0 CFU / mL。人工接种的苹果汁中SRCA的检出限为7.1×10 1和7.1×10 0通过分别观察白色沉淀物和荧光的CFU / mL。与传统的PCR方法相比，SRCA的灵敏度至少高出100倍，检测下限至少低100倍。七十样品进行了调查A.耐热菌污染，以及与那些由传统的微生物的培养方法相比，结果显示100％的灵敏度，97.01％的特异性，和97.14％的准确度。总体而言，该方法是视觉和快速检测酸曲霉的潜在有用工具。
更新日期：2020-04-06
中文翻译：
快速和目视检测苹果汁中酸脂环酸杆菌的咸化滚动圆扩增法的发展
建立了一种基于盐化滚环扩增（SRCA）的核酸等温扩增新方法，用于苹果汁中酸热脂环酸杆菌的快速直观检测。使用十四个酸曲霉菌株和44个非酸曲霉菌株来确认特异性。通过肉眼观察白色沉淀物，SRCA的灵敏度为4.5×10 1 CFU / mL，而通过荧光可视化为4.5×10 0 CFU / mL。人工接种的苹果汁中SRCA的检出限为7.1×10 1和7.1×10 0通过分别观察白色沉淀物和荧光的CFU / mL。与传统的PCR方法相比，SRCA的灵敏度至少高出100倍，检测下限至少低100倍。七十样品进行了调查A.耐热菌污染，以及与那些由传统的微生物的培养方法相比，结果显示100％的灵敏度，97.01％的特异性，和97.14％的准确度。总体而言，该方法是视觉和快速检测酸曲霉的潜在有用工具。
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