• Title/Summary/Keyword: absolute quantification

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A Recommendation System using Dynamic Profiles and Relative Quantification

  • Lee, Se-Il;Lee, Sang-Yong
    • International Journal of Fuzzy Logic and Intelligent Systems
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    • v.7 no.3
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    • pp.165-170
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    • 2007
  • Recommendation systems provide users with proper services using context information being input from many sensors occasionally under ubiquitous computing environment. But in case there isn't sufficient context information for service recommendation in spite of much context information, there can be problems of resulting in inexact result. In addition, in the quantification step to use context information, there are problems of classifying context information inexactly because of using an absolute classification course. In this paper, we solved the problem of lack of necessary context information for service recommendation by using dynamic profile information. We also improved the problem of absolute classification by using a relative classification of context information in quantification step. As the result of experiments, expectation preference degree was improved by 7.5% as compared with collaborative filtering methods using an absolute quantification method where context information of P2P mobile agent is used.

Development of the Droplet Digital PCR Method for the Detection and Quantification of Erwinia pyrifoliae

  • Lin, He;Seong Hwan, Kim;Jun Myoung, Yu
    • The Plant Pathology Journal
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    • v.39 no.1
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    • pp.141-148
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    • 2023
  • Black shoot blight disease caused by Erwinia pyrifoliae has serious impacts on quality and yield in pear production in Korea; therefore, rapid and accurate methods for its detection are needed. However, traditional detection methods require a great deal of time and fail to achieve absolute quantification. In the present study, we developed a droplet digital polymerase chain reaction (ddPCR) method for the detection and absolute quantification of E. pyrifoliae using a pair of species-specific primers. The detection range was 103-107 copies/ml (DNA templates) and cfu/ml (cell culture templates). This new method exhibited good linearity and repeatability and was validated by absolute quantification of E. pyrifoliae DNA copies from samples of artificially inoculated immature pear fruits. Here, we present the first study of ddPCR assay for the detection and quantification of E. pyrifoliae. This method has potential applications in epidemiology and for the early prediction of black shoot blight outbreaks.

High-accuracy quantitative principle of a new compact digital PCR equipment: Lab On An Array

  • Lee, Haeun;Lee, Cherl-Joon;Kim, Dong Hee;Cho, Chun-Sung;Shin, Wonseok;Han, Kyudong
    • Genomics & Informatics
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    • v.19 no.3
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    • pp.34.1-34.6
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    • 2021
  • Digital PCR (dPCR) is the third-generation PCR that enables real-time absolute quantification without reference materials. Recently, global diagnosis companies have developed new dPCR equipment. In line with the development, the Lab On An Array (LOAA) dPCR analyzer (Optolane) was launched last year. The LOAA dPCR is a semiconductor chip-based separation PCR type equipment. The LOAA dPCR includes Micro Electro Mechanical System that can be injected by partitioning the target gene into 56 to 20,000 wells. The amount of target gene per wells is digitized to 0 or 1 as the number of well gradually increases to 20,000 wells because its principle follows Poisson distribution, which allows the LOAA dPCR to perform precise absolute quantification. LOAA determined region of interest first prior to dPCR operation. To exclude invalid wells for the quantification, the LOAA dPCR has applied various filtering methods using brightness, slope, baseline, and noise filters. As the coronavirus disease 2019 has now spread around the world, needs for diagnostic equipment of point of care testing (POCT) are increasing. The LOAA dPCR is expected to be suitable for POCT diagnosis due to its compact size and high accuracy. Here, we describe the quantitative principle of the LOAA dPCR and suggest that it can be applied to various fields.

Selective Extraction and Quantification of Glutathione using Maleimide-Presenting Gold Nanoparticles

  • Oh, Hongseok;Lee, Jeongwook;Yeo, Woon-Seok
    • Bulletin of the Korean Chemical Society
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    • v.35 no.10
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    • pp.3047-3051
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    • 2014
  • In this paper, we describe a new method for the selective extraction and quantification of glutathione (GSH) using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and maleimide-presenting gold nanoparticles (Mal-AuNPs). Our strategy utilizes the Michael addition to selectively extract GSH, from chosen samples, onto the maleimide of Mal-AuNPs. After the extraction step, the GSH bound to the AuNPs was analyzed by MALDI-TOF MS in the presence of an internal standard which was prepared by reacting Mal-AuNPs with isotope-labeled GSH ($GSH^*$). The $GSH^*$ has the same structure as GSH but a higher molecular weight, and therefore, enables absolute quantification of GSH by comparing the mass signal intensities of the GSH- and $GSH^*$-conjugated alkanethiols. Our strategy was verified by analyzing GSH-spiked fetal bovine serum and NIH 3T3 cells.

Quantification Method of Kinesthetic Sense of Arm with Compensation for Individual Difference (개인차 보상을 고려한 팔운동감의 정량화 방법)

  • 정웅철;송재복;김용일
    • Journal of the Korean Society for Precision Engineering
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    • v.17 no.2
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    • pp.227-234
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    • 2000
  • The human sensibility caused by the motion of an object grasped by a human operator is defined as kinesthetic sense of arm. Due to nonlinearity and ambiguity of human sense, there is no absolute standard for quantification of kinesthetic sense. In this research, a so-called 2-dimensional arm motion generator is developed to present various mechanical impedance (i.e., stiffness or damping) characteristics to a human arm. The kinesthetic words representing arm kinesthetic sense are selected and then the subject's satisfaction levels on these words for given impedance values are measured and processed by the SD method and factor analysis. In addition, the quantification method using neural network is proposed to take into account the individual difference between the mean sensibility and each subject's sensibility. Through this proposed algorithm, the sensibility of human motion described qualitatively can be converted into engineering data ensuring objectivity, reproducibility, and universality.

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Quantification of Myocardial Perfusion and Function Using SPECT and PET (SPECT와 PET을 이용한 심장관류 및 기능의 정량화)

  • Lee, Jae-Sung
    • The Korean Journal of Nuclear Medicine
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    • v.39 no.2
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    • pp.75-81
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    • 2005
  • Myocardial perfusion and function can be quantified using SPECT and PET. There was controversy over the usefulness of the correction techniques for physical artifacts, such as photon attenuation and scatter, in the quantification of myocardial perfusion using SPECT. However, the cumulated results of many investigations have leaded the consensus on the usefulness of the correction procedures to improve the accuracy and specificity of the myocardial SPECT in the assessment of coronary artery diseases. Although the clinical value of the myocardial perfusion PET has not been preyed yet, the absolute myocardial blood flow and perfusion reserve values quantified using myocardial PET are employed in many basic investigations. In this paper, the methods for the quantitative myocardial SPECT and PET will be reviewed.

Epifluorescence Microscopy with Image Analysis as a Promising Method for Multispecies Biofilm Quantification

  • Ji Won Lee;So-Yeon Jeong;Tae Gwan Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.3
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    • pp.348-355
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    • 2023
  • Epifluorescence microscopy with image analysis was evaluated as a biofilm quantification method (i.e., quantification of surface area colonized by biofilms), in comparison with crystal violet (CV) staining. We performed different experiments to generate multispecies biofilms with natural and artificial bacterial assemblages. First, four species were inoculated daily in 16 different sequences to form biofilms (surface colonization, 0.1%-56.6%). Second, a 9-species assemblage was allowed to form biofilms under 10 acylase treatment episodes (33.8%-55.6%). The two methods comparably measured the quantitative variation in biofilms, exhibiting a strong positive relationship (R2 ≥ 0.7). Moreover, the two methods exhibited similar levels of variation coefficients. Finally, six synthetic and two natural consortia were allowed to form biofilms for 14 days, and their temporal dynamics were monitored. The two methods were comparable in quantifying four biofilms colonizing ≥18.7% (R2 ≥ 0.64), but not for the other biofilms colonizing ≤ 3.7% (R2 ≤ 0.25). In addition, the two methods exhibited comparable coefficients of variation in the four biofilms. Microscopy and CV staining comparably measured the quantitative variation of biofilms, exhibiting a strongly positive relationship, although microscopy cannot appropriately quantify the biofilms below the threshold colonization. Microscopy with image analysis is a promising approach for easily and rapidly estimating absolute quantity of multispecies biofilms.

Investigation on the Stability of Uric Acid and its Isotope (1,3-15N2) in Ammonium Hydroxide for the Absolute Quantification of Uric Acid in Human Serum

  • Lee, Sun Young;Kim, Kwonseong;Oh, Han Bin;Hong, Jongki;Kang, Dukjin
    • Mass Spectrometry Letters
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    • v.8 no.3
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    • pp.59-64
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    • 2017
  • In clinical diagnosis, it's well known that the abnormal level of uric acid (UA) in human body is implicated in diverse human diseases, for instance, chronic heart failure, gouty arthritis, diabetes, and so on. As a primary method, an isotope dilution mass spectrometry (IDMS) has been used to obtain the accurate quantity of UA in blood or serum and also develop the certificated reference material (CRM) so as to provide a SI-traceability to clinical laboratories. Due to the low solubility of UA in water, an ammonium hydroxide ($NH_4OH$) has been considered as a promising solvent to increase the solubility of UA that enables the preparation of both UA and its isotope standard solution for next IDMS-based absolute quantification. But, because of using this $NH_4OH$ solvent, it gives rise to the unwanted degradation of UA. In this study, we sought to optimize condition for the stability of UA in $NH_4OH$ solution by varying the mole ratios of UA to $NH_4OH$, followed by ID-LC-MRM analysis. In addition, we also inspected minutely the effect of the storage temperatures. Additionally, we also performed the quantitative analysis of UA in the KRISS serum certificated reference material (CRM, 111-01-02A) with diverse mixing ratios of UA to $NH_4OH$ and then compared those values to its certification value. Based on our experiments, adjusting the mole ratio of 1/2 ($UA/NH_4OH$) with the storage temperature of $-20^{\circ}C$ is an effective way to secure both the solubility and stability of UA in $NH_4OH$ solution for next IDMS-based quantification of UA in serum.

Comparison of microbial molecular diagnosis efficiency within unstable template metagenomic DNA samples between qRT-PCR and chip-based digital PCR platforms

  • Dongwan Kim;Junhyeon Jeon;Minseo Kim;Jinuk Jeong;Young Mok Heo;Dong-Geol Lee;Dong Keon Yon;Kyudong Han
    • Genomics & Informatics
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    • v.21 no.4
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    • pp.52.1-52.10
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    • 2023
  • Accurate and efficient microbial diagnosis is crucial for effective molecular diagnostics, especially in the field of human healthcare. The gold standard equipment widely employed for detecting specific microorganisms in molecular diagnosis is quantitative real-time polymerase chain reaction (qRT-PCR). However, its limitations in low metagenomic DNA yield samples necessitate exploring alternative approaches. Digital PCR, by quantifying the number of copies of the target sequence, provides absolute quantification results for the bacterial strain. In this study, we compared the diagnostic efficiency of qRT-PCR and digital PCR in detecting a particular bacterial strain (Staphylococcus aureus), focusing on skin-derived DNA samples. Experimentally, specific primer for S. aureus were designed at transcription elongation factor (greA) gene and the target amplicon were cloned and sequenced to validate efficiency of specificity to the greA gene of S. aureus. To quantify the absolute amount of microorganisms present on the skin, the variable region 5 (V5) of the 16S rRNA gene was used, and primers for S. aureus identification were used to relative their amount in the subject's skin. The findings demonstrate the absolute convenience and efficiency of digital PCR in microbial diagnostics. We suggest that the high sensitivity and precise quantification provided by digital PCR could be a promising tool for detecting specific microorganisms, especially in skin-derived DNA samples with low metagenomic DNA yields, and that further research and implementation is needed to improve medical practice and diagnosis.