• Title/Summary/Keyword: DNA-based Identification

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Identification and Phylogenetic Relationship of Dermatophytes Based on RFLP Analysis and Nucleotide Sequence of Internal Transcribed Spacer (ITS)1 in Nuclear Ribosome DNA (ITS-RFLP와 ITS1 염기서열 분석에 의한 피부사상균의 동정과 계통적 유연관계)

  • Choi, Yeon-Hwa;Lee, Yeong-Seon;Yoo, Jae-Il;Kim, Bong-Su
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.1
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    • pp.49-60
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    • 2000
  • ITSI-5.8S-ITSII rDNA region was amplified from the reference strains and clinical isolates with ITS1 and ITS4 primers. These primers amplified DNA fragments of 550 bp in Microsporum audouinii and Trichophyton violaceum, 700 bp in Microsporum gypseum, Trichophyton mentagrophytes, Trichophyton rubrum, and Trichophyton tonsurans, and 750 bp in Microsporum ferreugineum and Microsporum canis. The restriction enzyme patterns of PCR products digested with 13 restriction enzyme including PstI were distint among the genera, whereas identical in the same species. Examination of the ITS (Internal Transcribed Spacers)1 nucleotide sequence revealed that there was the genetic difference in each genera and species. Phylogenetic relationship among each species showed that the Trichophyton mentagrophytes was more closely related Trichophyton tonsurans than Trichophyton rubrum, and Microsporum gypseum was less related than Microsporum spp..

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Distribution of Dominant Bifidobacteria in the Intestinal Microflora of Korean Adults and Seniors, Identified by SDS-PAGE of Whole Cell Proteins and 16S rDNA Sequence Analysis

  • KIM TAE WOON;SONG HEE SUNG;KIM HAE YEONG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.388-394
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    • 2005
  • In order to investigate the distribution of dominant Bifidobacterium species in intestinal microflora of Korean adults and seniors, SDS-PAGE profiles of whole cell proteins were used for the identification of bifidobacteria. To confirm the reliability of SDS-PAGE, the Bifidobacterium species identified by SDS-PAGE of whole cell proteins were validated by using 16S rDNA sequencing analysis. The results of SDS­PAGE corresponded well with those determined by the analysis of 16S rDNA sequencing. Based on the analysis of SDS-PAGE patterns on unidentified fecal strains which showed positive in fructose-6-phosphate phosphoketolase activity, B. adolescentis, B. longum, and B. bifidum were identified in the feces of adults, and B. adolescentis, B. longum, B. bifidum, B. breve, and B. dentium were identified in those of seniors. In most of the fecal samples tested, the predominant Bifidobacterium species consisted of only a few species, and differences in the distribution and numbers of Bifidobacterium species were observed between adults and seniors. B. adolescentis and B. longum were found to be the most common species in feces of adults, but not in seniors. Accordingly, the distribution and abundance of bifidobacteria in the human intestinal microflora varied depending on the age of hosts.

Rapid differentiation of Hanwoo and Holstein meat using multiplex allele specific polymerase chain reaction protocols (Multiplex allele specific PCR 방법을 이용한 한우고기와 젖소고기의 신속한 판별)

  • Koh, Ba-Ra-Da
    • Korean Journal of Veterinary Research
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    • v.45 no.3
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    • pp.351-357
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    • 2005
  • Here I describe a multiplex allele specific PCR-based approach for the rapid detection between Hanwoo and Holstein meat associated with Melanocortin 1 receptor (MC1R) gene. Specific and universal oligonucleotide primers were used in combination to detect the presence of a single nucleotide polymorphism within the bovine MC1R DNA sequence. The presence of the bovine MC1R gene is indicated by the production of a single control PCR product, whilst positive samples generate an alternative smaller specific product over the same region. The mutations in MC1R104 codon revealed depending on the presence or absence of an indicative fragment amplified from the wild-type allele of this codon. As little as 0.39 ng and 1.56 ng of genomic DNA of Hanwoo and Holstein could be detected by MAS-PCR assay, respectively. This technique, which is widely used in human genetic screening, provides a reliable and sensitive result that has not been documented for the identification of bovine coat color. The MAS-PCR assay approach was proven to be useful in complementing routine beef DNA analysis for differentiation of these MC1R variants and it would facilitate the screening of deceiving sales of Holstein meat in the butcher shop.

Morphology of a Larval Atlantic Footballfish Himantolophus groenlandicus Reinhardt, 1837 (Lophiiformes: Himantolophidae) Identified by Complete Mitochondrial DNA (미토콘드리아 전장 유전체로 동정한 아귀목 Himantolophus groenlandicus 자어의 형태적 특징)

  • Choi, Hae-young;Jang, Yo-soon;Kim, Sung
    • Korean Journal of Ichthyology
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    • v.34 no.1
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    • pp.1-7
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    • 2022
  • A larva of the deep-sea angler fish, Himantolophus groenlandicus (2.2 mm BL), identified based on the complete mitochondrial DNA sequence, was collected at the surface of the western North Pacific. The postflexion stage larva had a round body, small teeth, incipient dorsal fin rays, eyes slightly recessed in the lower part, and melanophores on the gills and parietal and dorsal regions. These morphological features differ from a description of a larva reported as the same species with similar size (2.1 mm BL). The genetic and morphological information of our specimen should be useful for identifying larval H. groenlandicus.

Quantification of the ichthyotoxic raphidophyte Chattonella marina complex by applying a droplet digital PCR

  • Juhee, Min;Kwang Young, Kim
    • ALGAE
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    • v.37 no.4
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    • pp.281-291
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    • 2022
  • Quantifying the abundance of Chattonella species is necessary to effectively manage the threats from ichthyotoxic raphidophytes, which can cause large-scale mortality of aquacultured fish in temperate waters. The identification and cell counting of Chattonella species have been conducted primarily on living cells without fixation by light microscopy because routine fixatives do not retain their morphological features. Species belonging to the Chattonella marina complex, including C. marina and C. marina var. ovata, had high genetic similarities and the lack of clear morphological delimitations between the species. To estimate the abundance of C. marina complex in marine plankton samples, we developed a protocol based on the droplet digital polymerase chain reaction (ddPCR) assay, with C. marina complex-specific primers targeting the internal transcribed spacer (ITS) region of the rDNA. Cell abundance of the C. marina complex can be determined using the ITS copy number per cell, ranging from 25 ± 1 for C. marina to 112 ± 7 for C. marina var. ovata. There were no significant differences in ITS copies estimated by the ddPCR assay between environmental DNA samples from various localities spiked with the same number of cells of culture strains. This approach can be employed to improve the monitoring efficiency of various marine protists and to support the implementation of management for harmful algal blooms, which are difficult to analyze using microscopy alone.

Indirect Adaptive Control of Nonlinear Systems Using a EKF Learning Algorithm Based Wavelet Neural Network (확장 칼만 필터 학습 방법 기반 웨이블릿 신경 회로망을 이용한 비선형 시스템의 간접 적응 제어)

  • Kim Kyoung-Joo;Choi Yoon Ho;Park Jin Bae
    • Journal of the Korean Institute of Intelligent Systems
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    • v.15 no.6
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    • pp.720-729
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    • 2005
  • In this paper, we design the indirect adaptive controller using Wavelet Neural Network(WNN) for unknown nonlinear systems. The proposed indirect adaptive controller using WNN consists of identification model and controller. Here, the WNN is used in both Identification model and controller The WNN has advantage of indicating the location in both time and frequency simultaneously, and has faster convergence than MLPN and RBFN. There are several training methods for WNN, such as GD, GA, DNA, etc. In this paper, we present the Extended Kalman Filter(EKF) based training method. Although it is computationally complex, this algorithm updates parameters consistent with previous data and usually converges in a few iterations. Finally, ore illustrate the effectiveness of our method through computer simulations for the Buffing system and the one-link rigid robot manipulator. From the simulation results, we show that the indirect adaptive controller using the EKF method has better performance than the GD method.

Candida non albicans with a High Amphotericin B Resistance Pattern Causing Candidemia among Cancer Patients

  • Kalantar, Enayatollah;Assadi, Mojan;Pormazaheri, Helen;Hatami, Shiva;Barari, Maryam Agha;Asgari, Esfandiar;Mahmoudi, Elaheh;Kabir, Kourosh;Marashi, Seyed Mahmoud Amin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10933-10935
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    • 2015
  • Background: Many scientists have reported Candida species to be of great concern because of the high frequency that they colonize and infect human hosts, particularly cancer patients. Moreover, in the last decades Candida species have developed resistance to many antifungal agents. Based on this, we aimed to identify and determine the prevalence of Candida spp from blood culture bottles among cancer patients and their antifungal resistance pattern. Materials and Methods: From the blood culture bottles isolation and identification of the Candida spp were performed by conventional microbiological techniques. The in vitro antibiotic resistance pattern of the isolates was determined by CLSI guidelines. Genomic DNA was isolated and amplified. Each gene was separated by agar gel electrophoresis. Results: Identification of Candida spp was based on the presence of yeast cells in direct examination, culture and DNA extraction. Of the 68 blood samples collected during the study period (April 2013 to October 2013), five (7.35%) were positive for the presence of Candida spp, 2 (40%) of which were identified as Candida albicans and 3 (60%) were Candida non-albicans. Conclusions: High resistance to amphotricin B was observed among all the Candida non-albicans isolates. Regular investigations into antifungal resistance will help us to get an updated knowledge about their antibiotic resistance pattern which may help the physician in selecting the antibiotics for empirical therapy.

Development of PCR-based DNA markers for identification and detection of Trichoderma species associated with the green mold disease of oyster mushroom (느타리버섯 푸른곰팡이병에 관여하는 Trichoderma 속균의 동정 및 검출을 위한 PCR 기반 DNA 마커 개발)

  • Park, Myung Soo;Seo, Geon Sik;Ryu, Jae San;Kim, Min Kyung;Lee, Yong Kuk
    • Journal of Practical Agriculture & Fisheries Research
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    • v.24 no.3
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    • pp.5-14
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    • 2022
  • Trichoderma is known as pathogen caused serious green mold disease on commercial production. T. pleuroti and T. pleuroticola were common species in various mushroom media. Many strains of T. pleuroti, known as aggressive species causing major economic losses in Korea, showed benomyl resistance. Accurate identification and detection of Trichoderma species associated with oyster mushrooms is very important for disease control. We developed species-specific primers for T. pleuroticola, T. pleuroti, T. harzianum, and T. atroviride based on species-specific fragments isolated from amplified fragment length polymorphism analysis. PCR products corresponding to the predicted fragment of 500bp, 230bp, 180bp, and 410bp were amplified from T. pleuroticola, T. pleuroti, T. harzianum, and T. atroviride, respectively. Multiplex PCR assay using species-specific primers quickly and accurately identified and detected T. pleuroti from mushroom media in which various species co-exist. Our results can be useful for the effective control of mushroom disease.

Identification of ML106 Phase 1 Metabolites in Human Liver Microsomes Using High-Resolution Quadrupole-Orbitrap Mass Spectrometry

  • Jo, Jun Hyeon;Nam, WoongShik;Kim, Sunjoo;Lee, Doohyun;Min, Kyung Hoon;Lee, Taeho;Lee, Sangkyu
    • Mass Spectrometry Letters
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    • v.7 no.3
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    • pp.69-73
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    • 2016
  • High-resolution quadrupole-Orbitrap mass spectrometry (HRMS), with high-resolution (> 10,000 at full-width at half-maximum) and accurate mass (< 5 ppm deviation) capabilities, plays an important role in the structural elucidation of drug metabolites in the pharmaceutical industry. ML106, a derivative of imidazobenzimidazole, decreased melanin content and tyrosinase activity in a dose-dependent manner. Here, we investigated the phase 1 metabolic pathway of ML106 using HRMS in human liver microsomes (HLMs) and recombinant cDNA-expressed cytochrome P450 (CYP). After the incubation of ML106 with pooled HLMs and recombinant cDNA-expressed CYP in the presence of NADPH, five phase 1 metabolites, including three mono-hydroxylated metabolites (M1-3) and two di-hydroxylated metabolites (M4 and M5), were investigated. The metabolite structures were postulated by the elucidation of protonated mass spectra using HRMS. The CYP isoforms related to the hydroxylation of ML106 were studied after incubation with recombinant cDNA-expressed CYP. Here, we identified the phase 1 metabolic pathway of ML106 induced by CYP in HLMs.

Molecular Identification and Morphological Characteristics of the Longjaw Bonefish, Albula argentea Collected in Korea (한국산 여울멸의 분자동정과 형태적 특징)

  • Kim, Maeng Jin;Kim, Byeong Yeob;Song, Choon Bok
    • Korean Journal of Ichthyology
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    • v.31 no.4
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    • pp.249-254
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    • 2019
  • One specimen of Albula argentea (296.3 mm in standard length), belonging to the family Albulidae, was collected in a set net in the coastal waters of Jejudo Island, Korea. As the adult A. argentea has not been caught from Korean waters, we described its morphological characters based on the collected specimen and compared them with those of the previous reports on A. argentea and A. koreaus. As a result, the present specimen was characterized by having 68~73 pored lateral line scales (vs. 77~78 for A. koreana), 72~73 vertebrae pored lateral line scales (vs. 77~80) and none streak on the check (vs. yellowish streak). Molecular analysis using the mitochondrial cytochrome b DNA sequences was also conducted to confirm the correctness of species identification and taxonomic status of the sample.