该研究通过3管型微量MPN计数法(Mini Most Probable Number,mini-MPN)建立了一种快速定量检测食源性沙门氏菌的荧光定量环介导等温扩增(Quantitative Loop-Mediated Isothermal Amplification,qLAMP)方法。依据沙门菌属ttr基因设计了qL...该研究通过3管型微量MPN计数法(Mini Most Probable Number,mini-MPN)建立了一种快速定量检测食源性沙门氏菌的荧光定量环介导等温扩增(Quantitative Loop-Mediated Isothermal Amplification,qLAMP)方法。依据沙门菌属ttr基因设计了qLAMP和荧光定量PCR(Quantitative Polymerase Chain Reaction,qPCR)引物,结合5 h BPW增菌和MPN计数法建立了mini-MPN-qLAMP沙门氏菌快速定量检测方法;使用两种人工污染样品对mini-MPN-qLAMP法进行验证,使用Bland-Altman分析比较不同检测方法检测结果的一致性。结果表明,建立的qLAMP法与qPCR法反应特异性均良好,纯培养时qLAMP法检出限为500 CFU/mL。通过Bland-Altman分析表明所建立的mini-MPN-qLAMP法在速冻乌米饭中检测结果与mini-MPN-qPCR、mini-MPN计数法、平板计数法相比均具有较高的一致性,r^(2)≥0.994,检出限为-0.44 lg MPN/mL;而在速冻鸡胸肉中该法检测结果与mini-MPN-qPCR结果一致性最佳,r^(2)=0.990,检出限为-0.64 lg MPN/mL。肉制品中腐败杂菌会影响mini-MPN计数和平板计数结果,mini-MPN-qLAMP可排除肉制品中腐败杂菌对检测结果的影响。该研究所建立的mini-MPN-qLAMP法简单易行,准确度高,可用于食品中沙门氏菌的快速定量检测。展开更多
Background: It is important to achieve the definitive pathogen identification in hospital-acquired pneumonia (HAP), but the traditional culture results always delay the target antibiotic therapy. We assessed the me...Background: It is important to achieve the definitive pathogen identification in hospital-acquired pneumonia (HAP), but the traditional culture results always delay the target antibiotic therapy. We assessed the method called quantitative loop-mediated isothermal amplification (qLAMP) as a new implement for steering of the antibiotic decision-making in HAP. Methods: Totally, 76 respiratory tract aspiration samples were prospectively collected from 60 HAP patients. DNA was isolated from these samples. Specific DNA fragments for identifying 11 pneumonia-related bacteria were amplified by qLAMP assay. Culture results of these patients were compared with the qLAMP results. Clinical data and treatment strategies were analyzed to evaluate the effects of qLAMP results on clinical data. McNemar test and Fisher's exact test were used for statistical analysis. Results: The detection of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Stenotrophomonas maltophilia, Streptococcus pneumonia, and Acinetobacter baumannii by qLAMP was consistent with sputum culture (P 〉 0.05). The qLAMP results of 4 samples for Haemophilus influenzae, Legionella pneumophila, or Mvcoplasma pneumonia (MP) were inconsistent with culture results; however, clinical data revealed that the qLAMP results were all reliable except 1 MP positive sample due to the lack of specific species identified in the final diagnosis. The improvement of clinical condition was more significant (P 〈 0.001) in patients with pathogen target-driven therapy based on qLAMP results than those with empirical therapy. Conclusion: qLAMP is a more promising method for detection of pathogens in an early, rapid, sensitive, and specific manner than culture.展开更多
A model-flee compound controller design method is proposed to achieve the wide frequency bandwidth requirement of flight simulators. The method based on quantitative feedback theory, acquires system uncertainty under ...A model-flee compound controller design method is proposed to achieve the wide frequency bandwidth requirement of flight simulators. The method based on quantitative feedback theory, acquires system uncertainty under different working conditions through closed-loop identification with power spectrum estimation. Then in controller designing, it makes a trade, off between the strict requirements for magnitude-frequency characteristics and those for phase-frequency characteristics of flight simulators, by converting the indices of magnitude-frequency characteristics of flight simulators into quantitative feedback theory-based tracking specification bounds and using feedforward controller to attain the required phase-flequency characteristics. Simulation and experimental results indicate that, when used to design inner flame controller of flight simulator, the proposed method can fulfill the requirements for wide frequency bandwidth indices. Compared with other controller design methods, it has the property of model-free and transparency.展开更多
文摘该研究通过3管型微量MPN计数法(Mini Most Probable Number,mini-MPN)建立了一种快速定量检测食源性沙门氏菌的荧光定量环介导等温扩增(Quantitative Loop-Mediated Isothermal Amplification,qLAMP)方法。依据沙门菌属ttr基因设计了qLAMP和荧光定量PCR(Quantitative Polymerase Chain Reaction,qPCR)引物,结合5 h BPW增菌和MPN计数法建立了mini-MPN-qLAMP沙门氏菌快速定量检测方法;使用两种人工污染样品对mini-MPN-qLAMP法进行验证,使用Bland-Altman分析比较不同检测方法检测结果的一致性。结果表明,建立的qLAMP法与qPCR法反应特异性均良好,纯培养时qLAMP法检出限为500 CFU/mL。通过Bland-Altman分析表明所建立的mini-MPN-qLAMP法在速冻乌米饭中检测结果与mini-MPN-qPCR、mini-MPN计数法、平板计数法相比均具有较高的一致性,r^(2)≥0.994,检出限为-0.44 lg MPN/mL;而在速冻鸡胸肉中该法检测结果与mini-MPN-qPCR结果一致性最佳,r^(2)=0.990,检出限为-0.64 lg MPN/mL。肉制品中腐败杂菌会影响mini-MPN计数和平板计数结果,mini-MPN-qLAMP可排除肉制品中腐败杂菌对检测结果的影响。该研究所建立的mini-MPN-qLAMP法简单易行,准确度高,可用于食品中沙门氏菌的快速定量检测。
文摘Background: It is important to achieve the definitive pathogen identification in hospital-acquired pneumonia (HAP), but the traditional culture results always delay the target antibiotic therapy. We assessed the method called quantitative loop-mediated isothermal amplification (qLAMP) as a new implement for steering of the antibiotic decision-making in HAP. Methods: Totally, 76 respiratory tract aspiration samples were prospectively collected from 60 HAP patients. DNA was isolated from these samples. Specific DNA fragments for identifying 11 pneumonia-related bacteria were amplified by qLAMP assay. Culture results of these patients were compared with the qLAMP results. Clinical data and treatment strategies were analyzed to evaluate the effects of qLAMP results on clinical data. McNemar test and Fisher's exact test were used for statistical analysis. Results: The detection of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Stenotrophomonas maltophilia, Streptococcus pneumonia, and Acinetobacter baumannii by qLAMP was consistent with sputum culture (P 〉 0.05). The qLAMP results of 4 samples for Haemophilus influenzae, Legionella pneumophila, or Mvcoplasma pneumonia (MP) were inconsistent with culture results; however, clinical data revealed that the qLAMP results were all reliable except 1 MP positive sample due to the lack of specific species identified in the final diagnosis. The improvement of clinical condition was more significant (P 〈 0.001) in patients with pathogen target-driven therapy based on qLAMP results than those with empirical therapy. Conclusion: qLAMP is a more promising method for detection of pathogens in an early, rapid, sensitive, and specific manner than culture.
文摘A model-flee compound controller design method is proposed to achieve the wide frequency bandwidth requirement of flight simulators. The method based on quantitative feedback theory, acquires system uncertainty under different working conditions through closed-loop identification with power spectrum estimation. Then in controller designing, it makes a trade, off between the strict requirements for magnitude-frequency characteristics and those for phase-frequency characteristics of flight simulators, by converting the indices of magnitude-frequency characteristics of flight simulators into quantitative feedback theory-based tracking specification bounds and using feedforward controller to attain the required phase-flequency characteristics. Simulation and experimental results indicate that, when used to design inner flame controller of flight simulator, the proposed method can fulfill the requirements for wide frequency bandwidth indices. Compared with other controller design methods, it has the property of model-free and transparency.