Paenibacillus larvae is a gram-positive, spore-forming bacterium that is etiological agent for american foulbrood disease (AFB), which is the most severe disease in honey bee. To detect P. larvae from infected honeybee-comb or larvae, polyclonal antibody against whole bacterium was produced from guineapig and its specificity was evaluated. After optimization of ELISA-based detection system using these antibodies, a number of different P. larvae strains were analysed. Polyclonal antibody against P. larvae ATCC 25747 showed high affinity to most strains of P. larvae including P. larvae. strain ATCC 9545 (type strain), ATCC 25747 and other korean strain, SJl5 but exhibited no cross-reaction with other bacterial species. Additionally, this type of ELISA system was used for the detection of AFB in field-application The results have shown that this antibody could be useful for the rapid identification and monitoring of P. larvae in honeybee-comb.
Journal of the Korean Society of Environmental Restoration Technology
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v.17
no.1
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pp.111-122
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2014
The provided habitat of many services from natural capital is important. But because most ecosystem services tools qualitatively evaluated biodiversity or habitat quality, this study quantitatively analyzed those aspects using the species distribution model (MaxEnt). This study used location point data of the goat(Naemorhedus caudatus), marten(Martes flavigula), leopard cat(Prionailurus bengalensis), flying squirrel(Pteromys volans aluco) and otter(Lutra lutra) from the 3rd National Ecosystem Survey. Input data utilized DEM, landcover classification maps, Forest-types map and digital topographic maps. This study generated the MaxEnt model, randomly setting 70% of the presences as training data, with the remaining 30% used as test data, and ran five cross-validated replicates for each model. The threshold indicating maximum training sensitivity plus specificity was considered as a more robust approach, so this study used it to conduct the distribution into presence(1)-absence(0) predictions and totalled up a value of 5 times for uncertainty reduction. The test data's ROC curve of endangered mammals was as follows: growing down goat(0.896), otter(0.857), flying squirrel(0.738), marten(0.725), and leopard cat(0.629). This study was divided into two groups based on habitat: the first group consisted of the goat, marten, leopard cat and flying squirrel in the forest; and the second group consisted of the otter in the river. More than 60 percent of endangered mammals' distribution probability were 56.9% in the forest and 12.7% in the river. A future study is needed to conduct other species' distribution modeling exclusive of mammals and to develop a collection method of field survey data.
The selection of meaningful clinical tests and its reference values from a high-dimensional clinical data with imbalanced class distribution, one class is represented by a large number of examples while the other is represented by only a few, is an important issue for differential diagnosis between similar diseases, but difficult. For this purpose, this study introduces methods based on the concepts of both discernibility matrix and function in rough set theory (RST) with two discretization approaches, equal width and frequency discretization. Here these discretization approaches are used to define the reference values for clinical tests, and the discernibility matrix and function are used to extract a subset of significant clinical tests from the translated nominal attribute values. To show its applicability in the differential diagnosis problem, we have applied it to extract the significant clinical tests and its reference values between normal (N = 351) and abnormal group (N = 101) with either cholecystitis or cholelithiasis disease. In addition, we investigated not only the selected significant clinical tests and the variations of its reference values, but also the average predictive accuracies on four evaluation criteria, i.e., accuracy, sensitivity, specificity, and geometric mean, during l0-fold cross validation. From the experimental results, we confirmed that two discretization approaches based rough set approximation methods with relative frequency give better results than those with absolute frequency, in the evaluation criteria (i.e., average geometric mean). Thus it shows that the prediction model using relative frequency can be used effectively in classification and prediction problems of the clinical data with imbalanced class distribution.
The Journal of the Korean Society for Microbiology
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v.22
no.3
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pp.197-208
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1987
Chlamydia trachomatis has now shown that this interesting intracellular parasite is a cause of nongonococcal urethritis, infantile pneumonia, pelvic inflammatory disease and epididymitis, in addition to lymphogranuloma venerum and inclusion conjunctivitis. There are several diagnostic methods for C. trachomatis, but the method using monoclonal antibody is the most sensitive and specific. The hybride cell were prepared by fusion of myeloma cell($P_3X_{63}\;Ag_8{\cdot}V_{653}$) of mouse and lymphocyte of mouse(BALB/c) that were immunized with formalin killed C. trachomatis serotype D. The cell mixtures after fusion were dispensed into 640 wells of the 96 well culture plates and continuously cultured in HAT medium for 2 weeks. The supernatants of culture media in 83(13%) wells were reacted with C. trachomatis, which were determined by enzyme-linked immunosorbent assay in 96 well microplate. The clones that secreted antibody to C. trachomatis were cloned by limiting dilution. Only six monoclones secreted antibody to C. trachomatis. The antibody titer of ascitic fluid that collected from same BALB/c mice bearing hybridoma cells was above 1:100,000. These monoclonal antibodies that were IgG reacted with elementary and reticulate bodies of all serotypes(Ba, D, E, F, G, H, J and LGV type-I) using ELISA and indirect immunofluorescence stain, but there were no cross reaction with other bacteria(coagulase negative Staphylococcus, Proteus and E. coli). We concluded these six monoclones secreted the same monoclonal antibody to C. trachomatis. The sensitivity and specificity of the monoclonal antibody compared with Microtrak(confirmatory test of C. trachomatis, Syva) was 100%, respectively.
Background: Peste des petits ruminants (PPR) is an infectious disease caused by the peste des petits ruminants virus (PPRV) that mainly produces respiratory symptoms in affected animals, resulting in great losses in the world's agriculture industry every year. Single-domain variable heavy chain (VHH) antibody fragments, also referred to as nanobodies, have high expression yields and other advantages including ease of purification and high solubility. Objectives: The purpose of this study is to obtain a single-domain antibody with good reactivity and high specificity against PPRV. Methods: A VHH cDNA library was established by immunizing camels with PPRV vaccine, and the capacity and diversity of the library were examined. Four PPRV VHHs were selected, and the biological activity and antigen-binding capacity of the four VHHs were identified by western blot, indirect immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) analyses. ELISA was used to identify whether the four VHHs were specific for PPRV, and VHH neutralization tests were carried out. ELISA and western blot analyses were used to identify which PPRV protein was targeted by VHH2. Results: The PPRV cDNA library was constructed successfully. The library capacity was greater than 2.0 × 106 cfu/mL, and the inserted fragment size was approximately 400 bp to 2000 bp. The average length of the cDNA library fragment was about 1000 bp, and the recombination rate was approximately 100%. Four single-domain antibody sequences were selected, and proteins expressed in the supernatant were obtained. The four VHHs were shown to have biological activity, close affinity to PPRV, and no cross-reaction with common sheep diseases. All four VHHs had neutralization activity, and VHH2 was specific to the PPRV M protein. Conclusions: The results of this preliminary research of PPRV VHHs showed that four screened VHH antibodies could be useful in future applications. This study provided new materials for inclusion in PPRV research.
Purpose: The aim of this study is to explore levels of suicidal ideation and identify subgroups of high suicidal risk among the depressed elderly in Korea. Methods: A descriptive cross-sectional design was adopted on secondary data from the 6th (1st year) Korean national health and nutrition examination survey (KNHANES). A total of 239 depressed elders aged 60 or over who participated in the KNHANES. The prevalence of suicidal ideation and its related factors, including sociodemographic, physical, psychological characteristics and quality of life (EQ-5D index) were examined. Descriptive statistics and a decision tree analysis were performed using the SPSS/WIN 23.0 and SPSS Modeler 14.2 programs. Results: Of the depressed elderly, 28.9% had suicidal ideation. Three groups with high suicidal ideation were identified. Predictive factors included perceived stress level, household income level, quality of life and restriction of activity. In the highest risk group were those depressed elderly with moderate and low levels of stress, less than .71 of EQ-5D index and restriction of activity, and 80.0% of these participants had suicidal ideation. The accuracy of the model was 80.8%, its sensitivity 85.9%, and its specificity 68.1%. Conclusion: Multi-dimensional intervention should be designed to decrease suicide among the depressed elderly, particularly focusing on subgroups with high risk factors. This research is expected to contribute itself to the policy design and solution building in the future as it suggests policy implications in preventing the suicide of the depressed elderly.
Artemisinin resistance (ART) has been confirmed in Greater Mekong Sub-region countries. Currently, C580Y mutation on Pfkelch13 gene is known as the molecular marker for the detection of ART. Rapid and accurate detection of ART in field study is essential to guide malaria containment and elimination interventions. A simple method for collection of malaria-infected blood is to spot the blood on filter paper and is fast and easy for transportation and storage in the field study. This study aims to evaluate LAMP-SNP assay for C580Y mutation detection by introducing an extra mismatched nucleotide at the 3' end of the FIP primer. The LAMP-SNP assay was performed in a water bath held at a temperature of 56℃ for 45 min. LAMP-SNP products were interpreted by both gel-electrophoresis and HNB-visualized changes in color. The method was then tested with 120 P. falciparum DNA from dried blood spot samples. In comparing the LAMP-SNP assay results with those from DNA sequencing of the clinical samples, the 2 results fully agreed to detect C580Y. The sensitivity and specificity of the LAMP-SNP assay showed 100%. There were no cross-reactions with other Plasmodium species and other Pfkelch13 mutations. The LAMP-SNP assay performed in this study was rapid, reliable, and useful in detecting artemisinin resistance in the field study.
Porphyromonas gingivalis (P. gingivalis) is a major bacterial pathogen that causes periodontitis, a chronic inflammatory disease of tissues around the teeth. Periodontitis is known to be related to other diseases, such as oral cancer, Alzheimer's disease, and rheumatism. Thus, a precise and sensitive test to detect P. gingivalis is necessary for the early diagnosis of periodontitis. The objective of this study was to optimize a rapid visual detection system for P. gingivalis. First, we performed a visual membrane immunoassay using 3,3',5,5'-tetramethylbenzidine (TMB; blue) and coating and detection antibodies that could bind to the host laboratory strain, ATCC 33277. Antibodies against the P. gingivalis surface adhesion molecules RgpB (arginine proteinase) and Kgp (lysine proteinase) were determined to be the most specific coating and detection antibodies, respectively. Using these two selected antibodies, the streptavidin-horseradish peroxidase (HRP) reaction was performed using a nitrocellulose membrane and visualized with a detection range of 103-105 bacterial cells/ml following incubation for 15 min. These selected conditions were applied to test other oral bacteria, and the results showed that P. gingivalis could be detected without cross-reactivity to other bacteria, including Streptococcus mutans and Escherichia fergusonii. Furthermore, three clinical strains of P. gingivalis, KCOM 2880, KCOM 2803, and KCOM 3190, were also recognized using this optimized enzyme immunoassay (EIA) system. To conclude, we established optimized conditions for P. gingivalis detection with specificity, accuracy, and sensitivity. These results could be utilized to manufacture economical and rapid detection kits for P. gingivalis.
Journal of Korean Society of Industrial and Systems Engineering
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v.44
no.4
/
pp.12-22
/
2021
This article suggests the machine learning model, i.e., classifier, for predicting the production quality of free-machining 303-series stainless steel(STS303) small rolling wire rods according to the operating condition of the manufacturing process. For the development of the classifier, manufacturing data for 37 operating variables were collected from the manufacturing execution system(MES) of Company S, and the 12 types of derived variables were generated based on literature review and interviews with field experts. This research was performed with data preprocessing, exploratory data analysis, feature selection, machine learning modeling, and the evaluation of alternative models. In the preprocessing stage, missing values and outliers are removed, and oversampling using SMOTE(Synthetic oversampling technique) to resolve data imbalance. Features are selected by variable importance of LASSO(Least absolute shrinkage and selection operator) regression, extreme gradient boosting(XGBoost), and random forest models. Finally, logistic regression, support vector machine(SVM), random forest, and XGBoost are developed as a classifier to predict the adequate or defective products with new operating conditions. The optimal hyper-parameters for each model are investigated by the grid search and random search methods based on k-fold cross-validation. As a result of the experiment, XGBoost showed relatively high predictive performance compared to other models with an accuracy of 0.9929, specificity of 0.9372, F1-score of 0.9963, and logarithmic loss of 0.0209. The classifier developed in this study is expected to improve productivity by enabling effective management of the manufacturing process for the STS303 small rolling wire rods.
Vibrio parahaemolyticus is recognized as one of the most important foodborne pathogens responsible for gastroenteritis in humans. The blaCARB-17 gene is an intrinsic β-lactamase gene and a novel species-specific genetic marker of V. parahaemolyticus. In this study, a loop-mediated isothermal amplification (LAMP) assay combined with a lateral flow dipstick (LFD) was developed targeting this blaCARB-17 gene. The specificity of LAMP-LFD was ascertained by detecting V. parahaemolyticus ATCC 17802 and seven other non-V. parahaemolyticus strains. Finally, the practicability of LAMP-LFD was confirmed by detection with V. parahaemolyticus-contaminated samples and natural food samples. The results showed that the optimized reaction parameters of LAMP are as follows: 2.4 mmol/l Mg2+, 0.96 mmol/l dNTPs, 4.8 U Bst DNA polymerase, and an 8:1 ratio of inner primer to outer primer, at 63℃ for 40 min. The optimized reaction time of the LFD assay is 60 min. Cross-reactivity analysis with the seven non-V. parahaemolyticus strains showed that LAMP-LFD was exclusively specific for V. parahaemolyticus. The detection limit of LAMP-LFD for V. parahaemolyticus genomic DNA was 2.1 × 10-4 ng/μl, corresponding to 630 fg/reaction and displaying a sensitivity that is 100-fold higher than that of conventional PCR. LAMP-LFD in a spiking study revealed a detection limit of approximately 6 CFU/ml, which was similar with conventional PCR. The developed LAMP-LFD specifically identified the 10 V. parahaemolyticus isolates from 30 seafood samples, suggesting that this LAMP-LFD may be a suitable diagnostic method for detecting V. parahaemolyticus in aquatic foods.
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