• Korean Journal of Veterinary Service
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• v.45 no.1
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• pp.19-30
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• 2022

In this study, a simple loop-mediated isothermal amplification (LAMP) combined with visual detection method (vLAMP) assay was developed for the rapid and specific detection of African swine fever virus (ASFV), overcoming the shortcomings of previously described LAMP assays that require additional detection steps or pose a cross-contamination risk. The assay results can be directly detected by the naked eye using hydroxynaphthol blue after incubation for 40 min at 62℃. The assay specifically amplified ASFV DNA and no other viral nucleic acids. The limit of detection of the assay was <50 DNA copies/reaction, which was ten times more sensitive than conventional polymerase chain reaction (cPCR) and comparable to real-time PCR (qPCR). For clinical evaluation, the ASFV detection rate of vLAMP was higher than cPCR and comparable to OIE-recommended qPCR, showing 100% concordance, with a κ value (95% confidence interval) of 1 (1.00~1.00). Considering the advantages of high sensitivity and specificity, no possibility for cross-contamination, and being able to be used as low-cost equipment, the developed vLAMP assay will be a valuable tool for detecting ASFV from clinical samples, even in resource-limited laboratories.

• Journal of the Korean Society for Precision Engineering
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• v.11 no.4
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• pp.114-125
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• 1994

Fatigue behavior and life prediction method were presented for themal-mechanical and isothermal low cycle fatigue of 12 Cr forged steel used for high temperature applications. In-phase and out-of-phase thermal-mechanical fatigue test from 350 .deg. C to 600 .deg. C and isothermal low cycle fatigue test at 600 .deg. C, 475 .deg. C, 350 .deg. C were conducted using smooth cylindrical hollow specimen under strain-control with total strain ranges from 0.006 to 0.015. The phase difference between temperature and strain in thermal-mechanical fatigue resulted in significantly shorter fatigue life for out-of-phase than for in-phase. Thermal-mechanical fatigue life predication was made by partitioning the strain ranges of the hysteresis loops and the results of isothermal low cycle fatigue tests which were performed under the combination of slow and fast strain rates. Predicted fatigue lives for out-of-phase using the strain range partitioning method showed an excellent agreement with the actual out-of-phase thermal-mechanical fatigue lives within a factor of 1.5. Conventional strain range partitioning method exhibited a poor accuracy in the prediction of in-phase range partitioning method in a conservative way. By the way life prediction of thermal-mechanical fatigue by Taira's equivalent temperature method and spanning fartor method showed good agreement within out-of-phase thermal-mechanical fatigue.

• The Plant Pathology Journal
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• v.39 no.4
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• pp.309-318
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• 2023

Huanglongbing (HLB) is one of the most destructive diseases in citrus, which imperils the sustainability of citriculture worldwide. The presumed causal agent of HLB, 'Candidatus Liberibacter asiaticus' (CLas) is a non-culturable phloem-limited α-proteobacterium transmitted by Asian citrus psyllids (ACP, Diaphorina citri Kuwayama). A widely adopted method for HLB diagnosis is based on quantitative real-time polymerase chain reaction (qPCR). Although HLB diagnostic qPCR provides high sensitivity and good reproducibility, it is limited by time-consuming DNA preparation from plant tissue or ACP and the requirement of proper lab instruments including a thermal cycler to conduct qPCR. In an attempt to develop a quick assay that can be deployed in the field for CLas detection, we developed a real-time loop-mediated isothermal amplification (rt-LAMP) assay by targeting the CLas five copy nrdB gene. The rt-LAMP assay using various plant sample types and psyllids successfully detected the nrdB target as low as ~2.6 Log10 copies. Although the rt-LAMP assay was less sensitive than laboratory-based qPCR (detection limit ~10 copies), the data obtained with citrus leaf and bark and ACP showed that the rt-LAMP assay has >96% CLas detection rate, compared to that of laboratory-based qPCR. However, the CLas detection rate in fibrous roots was significantly decreased compared to qPCR due to low CLas titer in some root DNA sample. We also demonstrated that the rt-LAMP assay can be used with a crude leaf DNA extract which is fully deployable in the field for quick and reliable HLB screening.

• Journal of Microbiology and Biotechnology
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• v.33 no.8
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• pp.1101-1110
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• 2023

Streptococcus mutans is the primary causative agent of caries, which is one of the most common human diseases. Thus, rapid and early detection of cariogenic bacteria is critical for its prevention. This study investigated the combination of loop-mediated isothermal amplification (LAMP) and microfluid technology to quantitatively detect S. mutans. A low-cost, rapid microfluidic chip using LAMP technology was developed to amplify and detect bacteria at 2.2-2.2 × 10⁶ colony-forming units (CFU)/ml and its detection limits were compared to those of standard polymerase chain reaction. A visualization system was established to quantitatively determine the experimental results, and a functional relationship between the bacterial concentration and quantitative results was established. The detection limit of S. mutans using this microfluidic chip was 2.2 CFU/ml, which was lower than that of the standard approach. After quantification, the experimental results showed a good linear relationship with the concentration of S. mutans, thereby confirming the effectiveness and accuracy of the custom-made integrated LAMP microfluidic system for the detection of S. mutans. The microfluidic system described herein may represent a promising simple detection method for the specific and rapid testing of individuals at risk of caries.

• Journal of Food Hygiene and Safety
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• v.38 no.5
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• pp.347-355
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• 2023

In this study, polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) were compared in terms of their ability to detect shiga-toxin-producing Escherichia coli (STEC). Various foods were artificially inoculated with STEC to evaluate the limit of detection (LOD), limit of quantification (LOQ), sensitivity, specificity, and efficiency of PCR and LAMP. The LODs were ≤10⁴ and ≤10³ CFU/mL for PCR and LAMP, respectively. The LOQs did not differ between PCR and LAMP. However, of the four considered food types, the sensitivities differed by a maximum of 11.1% for seasoned meat and by a minimum of 8.1% for ground beef. LAMP had higher sensitivity than that of PCR and 100% specificity for all four food types. Therefore, LAMP is a reliable molecular method for detecting STEC as comparable to PCR assay, and its specificity and sensitivity are superior to those of PCR, depending on the food type.

• Tribology and Lubricants
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• v.40 no.1
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• pp.17-23
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• 2024

This study analyzes the thermal effects on the performance of an air foil thrust bearing (AFTB) using COMSOL Multiphysics to approximate actual bearing behavior under real conditions. An AFTB is a sliding-thrust bearing that uses air as a lubricant to support the axial load. The AFTB consists of top and bump foils and supports the rotating disk through the hydrodynamic pressure generated by the wedge effect from the inclined surface of the top foil and the elastic deformation of the bump foils, similar to a spring. The use of air as a lubricant has some advantages such as low friction loss and less heat generation, enabling air bearings to be widely used in high-speed rotating systems. However, even in AFTB, the effects of energy loss due to viscosity at high speeds, interface frictional heat, and thermal deformation of the foil caused by temperature increase cannot be ignored. Foil deformation derived from the thermal effect influences the minimum decay in film thickness and enhances the film pressure. For these reasons, performance analyses of isothermal AFTBs have shown few discrepancies with real bearing behavior. To account for this phenomenon, a thermal-fluid-structure analysis is conducted to describe the combined mechanics. Results show that the load capacity under the thermal effect is slightly higher than that obtained from isothermal analysis. In addition, the push and pull effects on the top foil and bump foil-free edges can be simulated. The differences between the isothermal and thermal behaviors are discussed.

• Journal of Adhesion and Interface
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• v.11 no.1
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• pp.15-25
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• 2010

In the present study, isothermal stabilization processes for rayon fabrics were performed at two relatively low temperatures $180^{\circ}C$ and $200^{\circ}C$ for a long period of time. The results of weight loss, dimensional shrinkage, X-ray diffraction and scanning electron microscopic observations studied with the rayon fabrics before and after the isothermal stabilization indicated that the chemical and physical changes of rayon precursor fibers proceeded continuously and slowly at the stabilization temperature below $200^{\circ}C$. And the pre-treatment with four different chemical compounds done prior to stabilization process influenced differently the characteristics of rayon fabrics. As a result, it was noticed that under the given stabilization conditions, $H_3PO_4$ and $Na_3PO_4$ played a role in catalyzing the stabilization reaction of rayon fabric whereas $NH_4Cl$ and $ZnCl_2$ played a role in delaying or retarding the reaction. $H_3PO_4$ showed the lowest percent weight loss of the fabric in the second stabilization conducted at $350^{\circ}C$. It was considered that phosphoric acid, which has a function of flame retardant, contributed to retarding somewhat the subsequent reaction even in the second stabilization step.

• Journal of the Korean Chemical Society
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• v.52 no.3
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• pp.273-280
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• 2008

Mycobacterium tuberculosis (MTB) remains a major worldwide public health problem. In recent years, the incidence of MTB has been rising. Rapid and reliable diagnosis of Mycobacterium tuberculosis is essential to initiate correct treatment, avoid severe complications, and prevent transmission. LAMP was used to develop a rapid and sensitive laboratory diagnostic system for the MTB. In this research, the loop-mediated isothermal amplification method (LAMP) that amplifies DNA with high specificity and rapidity at an isothermal condition was evaluated for rapid detection of MTB. Undiluted DNA (2.10 × 106 copy/mL), 10-1, 10-2, 10-3, 10-4, 10-5 and 10-6 (copy/mL) of MTB DNA were amplified by PCR and LAMP to determine the sensitivity of the assay. At results, the LAMP assay reported here has the advantages of rapid amplification, high sensitivity, and high specificity and will be useful for rapid and reliable clinical diagnosis of MTB in hospital clinical laboratory.

• Food Science of Animal Resources
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• v.33 no.5
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• pp.610-616
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• 2013

Loop-mediated isothermal amplification (LAMP) is an emerging detection technology for the amplification of DNA under isothermal conditions. The aim of this study was to develop a rapid and reliable LAMP technique for the detection of Cronobacter sakazakii in milk powder. In order to enhance the sensitivity and specificity, LAMP primers targeting outer membrane protein A (ompA) gene of C. sakazakii were designed using Explorer V4 software. Thirty seven C. sakazakii strains and 13 pathogenic microorganisms were used for comparative detection of C. sakazakii using polymerase chain reaction (PCR), real-time PCR, and LAMP. LAMP developed in this study could specifically detect C. sakazakii strains without cross-reactivity with other foodborne pathogens. LAMP products amplified from ompA gene of C. sakazakii were digested with with HhaI and NruI enzyme. The specificity of LAMP was confirmed by restriction fragment length polymorphism (RFLP) analysis. LAMP could detect C. sakazakii within 1 h without bacterial culture and its detection limit was as low as 1 CFU/mL C. sakazakii in milk. In the comparison of the sensitivity, LAMP showed 10,000- and 100-times higher detection limit than PCR or real-time PCR, respectively. Therefore, this study can conclude that LAMP is a rapid and reliable detection technique for C. sakazakii contaminated in powdered milk.

• The Korean Journal of Mycology
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• v.47 no.4
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• pp.437-441
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• 2019

Verticillium wilt disease is caused by a fungal plant pathogen Verticillium dahliae, which attacks commercial crops such as chrysanthemum. The conventional methods so far used to identify this fungal pathogen require high expertise and are time-consuming. Therefore, in this study, we developed an assay for the rapid and specific detection of V. dahliae infection using loop-mediated isothermal amplification (LAMP) method. For this assay, four primers for LAMP were designed for targeting cellulose-growth-specific protein partial mRNA gene in Verticillium dahliae. Under standard condition, the optimum reaction temperature for amplification is around 60 ℃ within 60 minutes. This LAMP assay was designed to amplify only present in V. dahliae. When this LAMP assay applied to the DNAs for four other soil-borne fungi and host plants, no amplification was detected. Therefore, this LAMP assay we developed for V. dahliae is expected to do detection at the early stage of its infection. The fast and reliable detection method will allow us to develop effective management system to monitor and control infection of this pathogen in chrysanthemum plant.