Vibrio anguillarum is an important bacterial pathogen of aquatic organisms and a significant problem in aquatic farming. The rapid detection and identification of V. anguillarum, and other pathogens that infect marine...Vibrio anguillarum is an important bacterial pathogen of aquatic organisms and a significant problem in aquatic farming. The rapid detection and identification of V. anguillarum, and other pathogens that infect marine organisms, is crucial to effective disease management. In this study, we developed a loop-mediated amplification （LAMP） assay to detect V. anguillarum in an hour in a single tube without the need for thermal cycling. Conserved regions of the metalloproteinase （empA） gene of V. anguillarum served as the targets for primer design. A fragment of the empA gene was amplified at 65℃ in the presence of the primer mixture and Bst DNA polymerase. In the optimized LAMP assay, 6.7 pg of V. anguillarum DNA could be detected. Six strains of V. anguillarum and 17 strains of non-V, anguillarum bacteria were used in this study to evaluate the species specificity of the primers. The six V. anguillarum strains gave a positive result in the LAMP assay. This method was also validated in V. anguillarum-infected fish. This LAMP method is more sensitive than PCR in the detection of V. anguillarum and shows good species specificity. The LAMP assay is therefore an effective method for the quick detection of V. anguillarum both in the laboratory and in the field.展开更多
This paper discusses the suitability of using TSA （thermoelastic stress analysis） as an advanced tool to detect damaged areas and highly stressed （hot spot） areas in structural components. Such components can be, ...This paper discusses the suitability of using TSA （thermoelastic stress analysis） as an advanced tool to detect damaged areas and highly stressed （hot spot） areas in structural components. Such components can be, for example, parts of large structural panels built of welded metallic or composite materials. Besides detecting hot spot areas, it is expected that stresses in these areas can be suitably quantified and processed in order to predict crack initiation and propagation due to in-service loads. The paper starts with references to selected review and application articles on the subject. Two simple laboratory experiments are presented which illustrate the quality of the results that can be achieved using TSA. In the first experiment, a stainless steel T-joint designed to model a welded structural component is analysed. The T-joint had a machine-notched crack-like flaw close to the component＇s weld toe. The qualitative and quantitative experimental results determined along four specified areas of the T-joint model showed that TSA can indeed be used as a tool to detect loaded cracks and hot spots in large metallic structures, and that stresses can be accurately evaluated. In the second experiment, a prismatic bar made of CFRE （carbon fibre-reinforced-epoxy） was tested to locate three subsurface areas of damage introduced beforehand into the component. Two of these inside damaged areas were detected to be 3.1 mm and 7.1 mm from the observed surface. The positive results achieved with the two lab experiments, along with a review of the selected research publications, indicate that TSA application can be extended to the real-world field of structural components. Topics to be addressed in this research field should have to do with components that work under random or quasi-cyclic service loading, problems where adiabatic conditions do not prevail, and reduction of the cost of infra-red cameras.展开更多
基金Supported by the National Basic Research Program of China (973 Program) (No. 2006CB101804)the General Administration of Quality Supervision,Inspection and Quarantine of the People's Republic of China (No. 2007IK167)National Department Public Benefit Research Foundation (No. 200803012)
文摘Vibrio anguillarum is an important bacterial pathogen of aquatic organisms and a significant problem in aquatic farming. The rapid detection and identification of V. anguillarum, and other pathogens that infect marine organisms, is crucial to effective disease management. In this study, we developed a loop-mediated amplification （LAMP） assay to detect V. anguillarum in an hour in a single tube without the need for thermal cycling. Conserved regions of the metalloproteinase （empA） gene of V. anguillarum served as the targets for primer design. A fragment of the empA gene was amplified at 65℃ in the presence of the primer mixture and Bst DNA polymerase. In the optimized LAMP assay, 6.7 pg of V. anguillarum DNA could be detected. Six strains of V. anguillarum and 17 strains of non-V, anguillarum bacteria were used in this study to evaluate the species specificity of the primers. The six V. anguillarum strains gave a positive result in the LAMP assay. This method was also validated in V. anguillarum-infected fish. This LAMP method is more sensitive than PCR in the detection of V. anguillarum and shows good species specificity. The LAMP assay is therefore an effective method for the quick detection of V. anguillarum both in the laboratory and in the field.
文摘This paper discusses the suitability of using TSA （thermoelastic stress analysis） as an advanced tool to detect damaged areas and highly stressed （hot spot） areas in structural components. Such components can be, for example, parts of large structural panels built of welded metallic or composite materials. Besides detecting hot spot areas, it is expected that stresses in these areas can be suitably quantified and processed in order to predict crack initiation and propagation due to in-service loads. The paper starts with references to selected review and application articles on the subject. Two simple laboratory experiments are presented which illustrate the quality of the results that can be achieved using TSA. In the first experiment, a stainless steel T-joint designed to model a welded structural component is analysed. The T-joint had a machine-notched crack-like flaw close to the component＇s weld toe. The qualitative and quantitative experimental results determined along four specified areas of the T-joint model showed that TSA can indeed be used as a tool to detect loaded cracks and hot spots in large metallic structures, and that stresses can be accurately evaluated. In the second experiment, a prismatic bar made of CFRE （carbon fibre-reinforced-epoxy） was tested to locate three subsurface areas of damage introduced beforehand into the component. Two of these inside damaged areas were detected to be 3.1 mm and 7.1 mm from the observed surface. The positive results achieved with the two lab experiments, along with a review of the selected research publications, indicate that TSA application can be extended to the real-world field of structural components. Topics to be addressed in this research field should have to do with components that work under random or quasi-cyclic service loading, problems where adiabatic conditions do not prevail, and reduction of the cost of infra-red cameras.
