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An algorithm for real time blood flow estimation of LDF (LDF의 실시간 혈류추정을 위한 알고리즘)

  • Kim, Jong-Weon;Ko, Han-Woo
    • Proceedings of the KOSOMBE Conference
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    • v.1998 no.11
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    • pp.78-79
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    • 1998
  • This paper describes a real time algorithm for blood flow estimation of LDF(laser Doppler flowmeter). Many algorithms for blood flow estimation are using power spectral density of Doppler signal by blood flow. In these research, the fast Fourier transformation is used to estimate power spectral density. This is a block processing procedure rather than real time processing. The algorithm in this paper used parametric spectral estimation. This has real time capability by estimation of AR(autoregressive) parameters sample by sample, and has smoothing power spectrum. Also, the frequency resolution is not limited by number of samples used to estimate AR parameter. Another advantage of this algorithm is that AR model enhance SNR.

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Comparison of quantitative detection of periodontal pathogens before and after scaling by real-time polymerase chain reaction

  • Kim, Young-Sun;Lee, Jung-Hwa;Lee, Young-Eun
    • Journal of Korean society of Dental Hygiene
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    • v.15 no.6
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    • pp.1063-1071
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    • 2015
  • Objectives: The purpose of the study is to investigate the quantitative detection of periodontal pathogens before and after scaling by real-time polymerase chain reaction. Methods: Participants were voluntarily recruited at D university, and saliva samples were extracted before and after scaling. Multiple real-time polymerase chain reactions were used to analyze characteristics and the amount of nine kinds of periodontal pathogens; Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Prevotella intermedia, Fusobacterium nucleatum, Parvimonas micra, Campylobacter rectus, and Eikenella corrodens. Results: After scaling, most periodontal pathogens except Eikenella corrodens were significantly decreased in all subjects(p<0.05). In addition, the percentage of microorganisms associated with disease, the microorganism risk index of periodontitis and the prevalence of red complex, orange complex, and Aggregatibacter actinomycetemcomitans was also significantly reduced after scaling(p<0.05). Conclusions: Scaling decreased in the amount of major periodontal pathogens and periodontitis prevalence rate.

PNA-mediated Real-Time PCR Clamping for Detection of EGFR Mutations

  • Choi, Jae-Jin;Cho, Min-Hey;Oh, Mi-Ae;Kim, Hyun-Sun;Kil, Min-Seock;Park, Hee-Kyung
    • Bulletin of the Korean Chemical Society
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    • v.31 no.12
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    • pp.3525-3529
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    • 2010
  • Tyrosine kinase inhibitors (TKIs) are currently used in the treatment of patients with advanced lung cancer. Recent studies on non-small cell lung cancer have shown that some patients carry somatic mutations in the epidermal growth factor receptor (EGFR) gene. Such mutations correlate with the effectiveness of certain TKIs. To detect a small amount of mutant EGFR among an abundance of wild-type EGFR, we have developed a highly sensitive and simple method using PNA-mediated real-time PCR clamping. The PNA-mediated real-time PCR clamping enables detection of EGFR mutants down to approximately 1% mutant -to- wild type. The total assay time was short as it required only 2.0 hr. Thus, PNA-mediated real-time PCR clamping can easily be applied to clinical samples for identification of DNA carrying EGFR mutations and also appear to be the best assay to detect somatic mutations.

Mining Information in Automated Relational Databases for Improving Reliability in Forest Products Manufacturing

  • Young, Timothy M.;Guess, Frank M.
    • International Journal of Reliability and Applications
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    • v.3 no.4
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    • pp.155-164
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    • 2002
  • This paper focuses on how modem data mining can be integrated with real-time relational databases and commercial data warehouses to improve reliability in real-time. An important Issue for many manufacturers is the development of relational databases that link key product attributes with real-time process parameters. Helpful data for key product attributes in manufacturing may be derived from destructive reliability testing. Destructive samples are taken at periodic time intervals during manufacturing, which might create a long time-gap between key product attributes and real-time process data. A case study is briefly summarized for the medium density fiberboard (MDF) industry. MDF is a wood composite that is used extensively by the home building and furniture manufacturing industries around the world. The cost of unacceptable MDF was as large as 5% to 10% of total manufacturing costs. Prevention can result In millions of US dollars saved by using better Information systems.

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Ultra Fast Real-Time PCR for Detection of Babesia gibsoni as Point of Care Test

  • Yang, Yong-Sung;Mun, Myung-Jun;Yun, Young-Min
    • Journal of Veterinary Clinics
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    • v.37 no.1
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    • pp.23-27
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    • 2020
  • Between May and November 2018, babesiosis was examined in 162 bloods samples obtained to an animal hospital in Jeju island for anemia and medical examination. Sixty-two of 162 (38.3%) were positive by PCR. The ultra fast real-time PCR test with blood directly analyzed without DNA extraction showed the same results. Accurate diagnosis, treatment and prognosis of babesiosis should be combined with clinical symptoms, blood tests, the babesia antibody test, and the PCR antigen test. Ultra fast real-time PCR, with these tests, is expected to be a point-of-care testing (POCT) for easy, fast and accurate diagnosis of babesiosis in the veterinary clinic.

Discovery of a new primer set for detection and quantification of Ilyonectria mors-panacis in soils for ginseng cultivation

  • Farh, Mohamed El-Agamy;Han, Jeong A.;Kim, Yeon-Ju;Kim, Jae Chun;Singh, Priyanka;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.43 no.1
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    • pp.1-9
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    • 2019
  • Background: Korean ginseng is an important cash crop in Asian countries. However, plant yield is reduced by pathogens. Among the Ilyonectria radicicola-species complex, I. mors-panacis is responsible for root-rot and replant failure of ginseng in Asia. The development of new methods to reveal the existence of the pathogen before cultivation is started is essential. Therefore, a quantitative real-time polymerase chain reaction method was developed to detect and quantify the pathogen in ginseng soils. Methods: In this study, a species-specific histone H3 primer set was developed for the quantification of I. mors-panacis. The primer set was used on DNA from other microbes to evaluate its sensitivity and selectivity for I. mors-panacis DNA. Sterilized soil samples artificially infected with the pathogen at different concentrations were used to evaluate the ability of the primer set to detect the pathogen population in the soil DNA. Finally, the pathogen was quantified in many natural soil samples. Results: The designed primer set was found to be sensitive and selective for I. mors-panacis DNA. In artificially infected sterilized soil samples, using quantitative real-time polymerase chain reaction the estimated amount of template was positively correlated with the pathogen concentration in soil samples ($R^2=0.95$), disease severity index ($R^2=0.99$), and colony-forming units ($R^2=0.87$). In natural soils, the pathogen was recorded in most fields producing bad yields at a range of $5.82{\pm}2.35pg/g$ to $892.34{\pm}103.70pg/g$ of soil. Conclusion: According to these results, the proposed primer set is applicable for estimating soil quality before ginseng cultivation. This will contribute to disease management and crop protection in the future.

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.

Surveillance of Acanthamoeba spp. and Naegleria fowleri in environmental water by using the duplex real-time PCR (Duplex real-time PCR을 이용한 수계 중 가시아메바와 파울러자유아메바 조사)

  • Kim, Min-jeong;Lee, Gyu-Cheol;Kim, Kunwoo;Lee, Hyunji;Kim, Min Young;Seo, Dae Keun;Lee, Jeong Yeob;Cho, Young-Cheol
    • Korean Journal of Microbiology
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    • v.54 no.2
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    • pp.98-104
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    • 2018
  • Naegleria fowleri and Acanthamoeba spp. are free-living amoebas that are widely distributed in natural environments. Although uncommon, infection with these protozoans can cause fatal disease in humans and animals. In this study, in order to select the appropriate method to survey Naegleria fowleri and Acanthamoeba spp. in water samples, four molecular biology techniques and one commercially available kit for real-time PCR were compared. The results indicated that the duplex real-time PCR was the most sensitive, and could be used to simultaneously detect two different free-living amoebas. Using the duplex real-time PCR approach, the two free-living amoebas were surveyed in three local streams in Daejeon, Republic of Korea. The concentrated free-living amoebas were inoculated onto non-nutrient agar plates which had been spread with heat-inactivated Escherichia coli and incubated for 5~7 days. After incubation, gDNA was extracted and used as the template for amplification by duplex real-time PCR. Acanthamoeba spp. and N. fowleri was detected from ten (83.3%) and two (16.6%) of the twelve samples, respectively. As these two free-living amoebas can be fatal, continuous surveillance is needed to track their distribution in the aquatic environment for the drinking water safety.

Species-specific Marker Development for Environmental DNA Assay of Endangered Bull-head Torrent Catfish, Liobagrus obesus (멸종위기어류 퉁사리의 환경 DNA 분석을 위한 종 특이 마커 개발)

  • Yun, Bong Han;Kim, Yong Hwi;Sung, Mu Sung;Han, Ho-Seop;Han, Jeong-Ho;Bang, In-Chul
    • Korean Journal of Ichthyology
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    • v.34 no.3
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    • pp.208-217
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    • 2022
  • We wanted to develop a real-time PCR assay capable of detecting Liobagrus obesus in environmental DNA (eDNA) extracted from freshwater samples using a pair of species-specific primers and probe for the endangered fish, L. obesus. The species-specific primers and probe were designed in consideration of single nucleotide polymorphisms between 65 species of freshwater fish living in the Republic of Korea within the cytochrome b (cytb) gene of mitochondrial DNA. The species-specific primers and probe, in the real-time PCR assay, showed high specificity as only the L. obesus genomic DNA (gDNA) was found to be positive in the specificity verification using 65 species gDNA of freshwater fish in the Republic of Korea. In addition, in the detection limit analysis using the serial dilution concentrations of L. obesus gDNA, it was found that it was possible to detect up to 0.2 pg, showing high sensitivity. Afterwards, using the species-specific primers and probe, real-time PCR assay was performed on freshwater samples obtained from 8 stations in the mid-upper basin of Geum River. As a result, the cytb gene of L. obesus was detected in total 5 stations including all 3 stations where this species was collected at the time of field survey. Therefore, the species-specific primers and probe developed in present study, and the real-time PCR assay using them, can accurately detect the cytb gene of L. obesus from eDNA samples, which can be utilized to monitor the existing habitats of this species and to discover potential new habitats.

A Real-Time PCR Assay for the Quantitative Detection of Ralstonia solanacearum in Horticultural Soil and Plant Tissues

  • Chen, Yun;Zhang, Wen-Zhi;Liu, Xin;Ma, Zhong-Hua;Li, Bo;Allen, Caitilyn;Guo, Jian-Hua
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.193-201
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    • 2010
  • A specific and rapid real-time PCR assay for detecting Ralstonia solanacearum in horticultural soil and plant tissues was developed in this study. The specific primers RSF/RSR were designed based on the upstream region of the UDP-3-O-acyl-GlcNAc deacetylase gene from R. solanacearum, and a PCR product of 159 bp was amplified specifically from 28 strains of R. solanacearum, which represent all genetically diverse AluI types and all 6 biovars, but not from any other nontarget species. The detection limit of $10^2\;CFU/g$ tomato stem and horticultural soil was achieved in this real-time PCR assay. The high sensitivity and specificity observed with field samples as well as with artificially infected samples suggested that this method might be a useful tool for detection and quantification of R. solanacearum in precise forecast and diagnosis.