Some phytopathogenic bacteria including Xanthomonas oryzae pv. oryzae (a rice leaf bacterial blight pathogen) and its virulent gene mutant XooM3105,Xanthomonas campestris pv. campestris, Erwinia carotovora subsp, ca...Some phytopathogenic bacteria including Xanthomonas oryzae pv. oryzae (a rice leaf bacterial blight pathogen) and its virulent gene mutant XooM3105,Xanthomonas campestris pv. campestris, Erwinia carotovora subsp, carotovora and so on, could release active oxygen such as O<sub>2</sub> and H<sub>2</sub>O<sub>2</sub> by themselves.Ca<sup>2+</sup>, NADPH and O<sub>2</sub> concentration regulated the response. The response is sensitive to heat and proteinase. These results show that the component releasing active oxygen in the bacterial plant pathogen may involve special protein and enzyme and be secreted to culture liquid. Various components of rice tissue can significantly induce the release of active oxygen in Xanthomonas oryzae pv. oryzae.展开更多
环介导等温扩增技术(loop-mediated isothermal amplification,LAMP)是快速检测食源性致病菌的常用方法。该文分别针对单增李斯特菌和金黄色葡萄球菌的特异性基因hlyA和nuc设计引物,通过优化反应条件,建立了一种可同时检测单增李斯特菌...环介导等温扩增技术(loop-mediated isothermal amplification,LAMP)是快速检测食源性致病菌的常用方法。该文分别针对单增李斯特菌和金黄色葡萄球菌的特异性基因hlyA和nuc设计引物,通过优化反应条件,建立了一种可同时检测单增李斯特菌和金黄色葡萄球菌的基于淬灭基团释放环介导等温扩增检测方法(detection of amplification by release of quenching-LAMP,DARQ-LAMP),并评估了该方法的特异性和灵敏度。结果表明该方法可在60 min内完成对单增李斯特菌和金黄色葡萄球菌的同时检测,检测限分别为7.3×10^(2)copies/mL和2.3×10^(2)copies/mL,特异性良好、灵敏度高,在实际样品检测中检测限为2.8×10^(4)CFU/mL和2.4×10^(4)CFU/mL。该双重DARQ-LAMP方法能同时检测2种食源性致病菌,缩短检测时间,提高了检测效率,为多重LAMP检测食源性致病菌的研究提供了新的思路。展开更多
文摘Some phytopathogenic bacteria including Xanthomonas oryzae pv. oryzae (a rice leaf bacterial blight pathogen) and its virulent gene mutant XooM3105,Xanthomonas campestris pv. campestris, Erwinia carotovora subsp, carotovora and so on, could release active oxygen such as O<sub>2</sub> and H<sub>2</sub>O<sub>2</sub> by themselves.Ca<sup>2+</sup>, NADPH and O<sub>2</sub> concentration regulated the response. The response is sensitive to heat and proteinase. These results show that the component releasing active oxygen in the bacterial plant pathogen may involve special protein and enzyme and be secreted to culture liquid. Various components of rice tissue can significantly induce the release of active oxygen in Xanthomonas oryzae pv. oryzae.
文摘环介导等温扩增技术(loop-mediated isothermal amplification,LAMP)是快速检测食源性致病菌的常用方法。该文分别针对单增李斯特菌和金黄色葡萄球菌的特异性基因hlyA和nuc设计引物,通过优化反应条件,建立了一种可同时检测单增李斯特菌和金黄色葡萄球菌的基于淬灭基团释放环介导等温扩增检测方法(detection of amplification by release of quenching-LAMP,DARQ-LAMP),并评估了该方法的特异性和灵敏度。结果表明该方法可在60 min内完成对单增李斯特菌和金黄色葡萄球菌的同时检测,检测限分别为7.3×10^(2)copies/mL和2.3×10^(2)copies/mL,特异性良好、灵敏度高,在实际样品检测中检测限为2.8×10^(4)CFU/mL和2.4×10^(4)CFU/mL。该双重DARQ-LAMP方法能同时检测2种食源性致病菌,缩短检测时间,提高了检测效率,为多重LAMP检测食源性致病菌的研究提供了新的思路。
文摘该研究针对单增李斯特氏菌的特异性基因hlyA设计引物,优化反应条件,建立实时荧光环介导等温扩增(realtime loop-mediated isothermal amplification,real-time LAMP)的检测方法,在此基础上,建立检测单增李斯特氏菌的基于淬灭基团释放环介导等温扩增检测方法(detection of amplification by release of quenching-LAMP,DARQ-LAMP),分析其特异性和灵敏度。结果表明,该方法的适宜反应条件为反应温度63℃、Bst DNA 3.0聚合酶0.32 U/μL、淬灭探针双链(quenching probe double chain,QPD)探针浓度10%、Mg2+浓度6 mmol/L;对单增李斯特菌的检测限为7.3×101copies/mL,灵敏度是普通聚合酶链式反应(polymerase chain reaction,PCR)的100倍。该方法效率高、特异性好、灵敏度高,为环介导等温扩增技术检测食源性致病菌的研究提供参考。