• Title/Summary/Keyword: water primer

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Genetic analysis of norovirueses in Busan (부산지역 노로바이러스의 유전적 분석)

  • Kim, Kwang-Il;Jin, Ji-Woong;Jeong, Hyun-Do
    • Journal of fish pathology
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    • v.24 no.3
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    • pp.255-268
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    • 2011
  • For detection of noroviruses (NVs), we compared various PCR primer sets based on reverse transcription nested PCR (RT-nested PCR) in the water samples from Dong brook in Busan, South Korea. We designed various new primer sets based on the most conserved sequences of the capsid protein gene that react with diverse NVs found in Korea. Designed primer sets (KG1F/KG1R and KG2F/KG2R, named as PNK) for the respective genogroups of NVs, genogroup I and II (GI and GII), were applied to detect NVs in the water samples from Dong brook concentrated with ultracentrifugation. In the application to the water samples, proportion of GI (76.47%) and GII (70.59%) in water samples of Dong brook in RT-nested PCR with the primer sets of this study. However, no significant differences of the proportion of the positive samples were not found between RT-nested PCRs with reported and newly designed primer sets. From the nucleotide sequencing, GI and GII of NVs present in Dong brook were appeared to be the members of 1/2/4/5/9/10 genotypes, and 3/4/5/11/13 genotypes respectively. Appeared genotype 4 of GII known as an one of main genotype found in patients of many Asian countries warned us to consider the risks of norovirus in aquatic environments in southern part of Korea.

Development of diagnostic method for human Astrovirus with rapid, specific and high sensitivity using loop-mediated isothermal amplification method

  • Lee, Jin-Young;Rho, Jae Young
    • Korean Journal of Agricultural Science
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    • v.47 no.1
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    • pp.173-182
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    • 2020
  • Human Astrovirus (HuAstV), known as a waterborne virus, is a group IV positive-sense single-stranded RNA that belongs to Astroviridae. The first outbreak of HuAstV was reported in England in 1975. HuAstV can exist not only among clinical patients but also in various water environments, such as water for agriculture and vegetables. For diagnosis of HuAstV from water samples, a polymerase chain reaction (PCR) system has been developed. However, the PCR-based diagnostic method has problems in field application, such as reaction time, sensitivity and specificity. For this reason, in this study we developed the loop-mediated isothermal amplification assay (LAMP) system, aimed specifically at HuAstV. Three prepared LAMP primer sets were tested by specificity, non-specificity and sensitivity; one LAMP primer set was selected with optimum reaction temperature. The developed LAMP primer set reaction conditions were confirmed at 62℃, and detection sensitivity was 1 fg/μL. In addition, restriction enzyme HaeIII (GG/CC) was introduced to confirm that the LAMP reaction was positive. As a result, selected LAMP primer set was 100 - 1000 times more specific, rapid, and sensitive than conventional-nested PCR methods. For verification of the developed LAMP assay, twenty samples of cDNA from groundwater samples were tested. We expect that the developed LAMP assay will be used to diagnose HuAstV from various samples.

Characteristics of Polyaniline Anti-Corrosive Coatings with Primer and Top Coating Resins (하도 및 상도 수지에 따른 폴리아닐린 방청도료의 특성)

  • Kim, Tae-Ok;Kong, Seung-Dae;Park, Jin-U
    • Journal of the Korean Applied Science and Technology
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    • v.24 no.4
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    • pp.399-409
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    • 2007
  • Characteristics of polyaniline anti-corrosive coatings with various primer coating resins(epoxy resin, urethane resin, and others) and top coating resins(epoxy and acrylic urethane resins) were investigated through adhesion, acid resistance, alkaline resistance, water resistance, and anti-corrosion tests. As a result, the anti-corrosive properties of the prepared coatings using polyaniline varied with the types of primer and top coating resins. In this condition, the properties of adhesion, chemical resistance, and water resistance were found to be very satisfactory when using emeraldine base (EB) of polyaniline blended with single-packaged urethane and acrylic urethane resins as the primer coatings, and using acrylic urethane resin as the top coatings. Also, the anti-corrosive function of these anti-corrosive coatings was well preserved for 1000 hr in the salt spray experiment.

Comparative Analysis of Microcystin during Water Treatment Process between Real-Time PCR and LC/MS (Real-Time PCR법과 LC/MS법을 이용한 수계중의 마이크로시스틴 검출방법 비교연구)

  • Park, Hong-Gi;Jung, Mi-Eun;Cha, Dong-Jin;Jung, Eun-Young;Bean, Jae-Hoon
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1201-1206
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    • 2010
  • We performed a comparative analysis using a Real-time PCR (Polymerase Chain Reaction) and LC/MS (Liquid-Chromatograph/Mass Spectrometer) method in order to detect microcystin in environmental sources. Among the three different primer sets tested for microcystin using three positive strains of Microcystis aeruginosa by Real-time PCR assay, only TOX2P/TOX2M primer pairs were able to detect Microcystis aeruginosa. According to the results of a survey carried out from June 2009 to September 2009, 11 out of 11 (100%) raw water samples were were found to have microcystin when the Real-Time PCR and LC/MS method was used, with total microcystin concentration ranging from 5.98~219.0 ${\mu}g/l$. A microcystin removal treatment process was used to ensure entire removal, by passing it through a BAC filtration step. It was concluded that real-time PCR assay can be used to estimate micrucystin detection more rapidly and easily than the LC/MS method.

BOND STRENGTH BETWEEN COBALT-CHROMIUM ALLOY AND DENTURE BASE RESIN ACCORDING TO ADHESIVE PRIMERS (금속표면처리제에 따른 코발트-크롬 합금과 의치상용 레진의 결합강도)

  • Park, Jong-Il;Kwon, Ju-Hong;Lee, Hae-Hyeung;Cho, Hay-Won
    • The Journal of Korean Academy of Prosthodontics
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    • v.38 no.2
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    • pp.160-168
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    • 2000
  • This study evaluated the effects of four adhesive metal primers on the shear bond strength of a heat curing denture base resin(Lucitone 199) to cobalt-chromium alloy(Biosil-f). The adhesive metal primers were Cesead Opaque Primer, Metal Primer, MR Bond, and Super-Bond liquid. The metal surface primed or nonprimed was filled with the heat-curing methyl methacrylate resin. The specimens were stored in water at $37^{\circ}C$ for 24 hours and the alternately immersed in water bath at $5^{\circ}C\;and\;55^{\circ}C$ for up to 2,000 thermal cycles. Shear bond strengths were measured using UTM at a crosshead speed of 0.5mm/min. Failure surface were examined under magnifying glasses. All the primers examined improved the shear bond strength between denture base resin and cobalt-chromium alloy compared with nonprimed specimens before thermal cycling. The bond strength of Cesead Opaque Primer was greatest. And after 2,000 thermal cycles, the bond strengths between resin and cobalt-chromium alloy were decreased but the difference between thermal cycling 0 and 2,000 at Cesead Opaque primer and Metal Primer were not significant. This study indicated that Cesead Opaque Primer & Metal Primer is effective primers to obtain higher bond strength between heat cured denture base resin and cobalt-chromium alloy.

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Molecular Detection of Harmful Dinoflagellates (Dinophyceae) in Ballast Water (선박평형 수 내 유해 와편모조류(Dinophyceae)의 분자생물학적 검출)

  • Park, Tae-Gyu;Kim, Sung-Yeon
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.15 no.1
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    • pp.36-40
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    • 2010
  • Ballast water has been known as a major vector for global dispersal of toxic dinoflagellates and other microalgae. In this study, biodiversity in ships’ ballast water was examined using a dinoflagellate-oriented PCR primer set and species-specific real-time PCR. While motile dinoflagellates could be observe at very low cell densities by light microscopy,a wide range of dinoflagellate taxa including parasitic and phototrophic pico-dinoflagellates as well as harmful species to marine fish/shellfish was detected when techniques for cloning/sequencing of SSU rDNA of sample cells were used. Present result suggests that molecular methods including species-specific PCR primers may offer rapid and accurate detection of invasive species in ballast water.

Detection of Toxigenicity of Cyanobacteria by Molecular Method (분자생물학적 방법에 의한 남조류의 독성 생성능의 확인)

  • Lee, Kyung-Lak;Jheong, Weon-Hwa;Kim, Jong-Min;Kim, Han-Soon
    • Korean Journal of Ecology and Environment
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    • v.40 no.1
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    • pp.149-154
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    • 2007
  • In the present study, we performed the PCR assay using TOX2P/TOX2M primer targeting a specific region within mcyB gene to identify potential microcystin-producing cyanobacteria. TOX2P/TOX2M primer set was effective in amplifying mcy gene in the field samples containing Microcystis spp. of 1,000 cells per mL. Moreover, the results from the PCR assay agreed with those of the ELISA analysis. Consequently, this study demonstrated that TOX2P/TOX2M primer set can be used as a genetic probe for the early detection of cyanobacterial toxigenicity in Korean water bodies.

EFFECT OF THERMOCYCLING ON BONDING OF COMPOSITE RESIN AFTER DENTIN SURFACE TREATMENTS (상아질 표면처리후 복합레진 수복시 온도변화가 결합에 미치는 영향)

  • Lee, Yong-Woo;Son, Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.16 no.2
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    • pp.126-142
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    • 1991
  • The purpose of this study was to evaluate the effect of various dentin surface treatments on shear bond strength, microhardness and fracture mode before and after thermocycling. Recently extracted 75 human molars were used. The teeth were sagittal sectioned faciolingually to obtain 150 specimens. They were randomly divided into six groups. Mesial and distal dentinal surfaces of specimens were exposed by grinding and treated respectively with GC-DENTIN CONDITIONER. 10-3 solution of 4-Meta, Cleansar and Primer of GLUMA, Scotchprep of Scotchbond 2, DENTIN CONDITIONER and PRIMER A, B of ALL BOND according to the manufacturers directions. Specimens of one group were not treated. Adhesive agent of Scotchbond 2, were applied and cured on the treated dentin surfaces. After P-50 were cured on them, specimens were stored in 31c water for 24 hours before shear bond strength measurement Shear bond strength was measured in 10 specimens of each group. 10 specimens of each group were thermocycled in $20^{\circ}C$, $60^{\circ}C$,$20^{\circ}C$, $4^{\circ}C$, $20^{\circ}C$ water in order, for 30 seconds respectively, 100 times a day for 7 days. After thermocycling shear bond strength was measured. Microhardness was checked on treated dentin surface and fractured dentin surface in 10 specimens respectievly. Francture modes were observed with SEM The following results were obtained. 1. Before thermocycling. shear bond strengths in the specimens treated with DENTIN CONDITIONER and PRIMER A, B of ALL BOND were significantly higher than those in other specimens(P<0.01). 2. After thermocycling. shear bond strengths in the specimens treated with Cleanser and Primer of GLUMA, Scotchprep of Scotchbond 2 and DENTIN CONDITIONER and PRIMER A, B of AIL BOND were significantly higher than those in specimens not: treated, treated with GC-DENTIN CONDITIONER and 10-3 solution of 4-Meta(P<0.01). Shear bond strengths in the specimens treated with GC-DENTIN CONDITIONER and PRIMER A, B of ALL BOND were significantly higher than those in other specimens except those treated with Scotchprep of Srotchbond 2(P<0.01). 3. Shear bond strengths after thermocycling were reduced in the specimens not treated, treated with GC-DENTIN CONDITIONER and 10-3 solution of 4-Meta and were increased in the specimens treated with Cleanser and Primer of GLUMA, Scotchprep of Scotchbond 2, without significance, compared with those before thermocycling. In the specimens treated with DENTIN CONDITIONER and PRIMER A, B of ALL BOND, shear bond strengths after thermocycling were significantly increased, compared with those before thermocycling(P<0.01). 4. Microhardnesses in the fractured surfaces after shear bond strength measurement were significantly increased in the specimens treated with 10-3 solution of 4-Meta and significantly decreased in the specimens treated with DENTIN CONDITIONER and PRIMER A, B of ALL BOND, compared with those in the treated dentin surfaces(P<0.01). 5. In the specimens treated with Cleanser and Primer of GLUMA, Scotchprep of Scotchbond 2 and DENTIN CONDITIONER and PRIMER A, B of ALL BOND, cohesive fracture modes were observed more than adhesive fracture modes.

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Detection of Denitrifying Bacteria in Groundwater by PCR (PCR을 이용한 지하수 내의 탈질화 세균의 검출)

  • Shin, Kyu-Chul;Suh, Mi-Yeon;Han, Myung-Soo;Choi, Yong-Keel
    • Korean Journal of Environmental Biology
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    • v.19 no.4
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    • pp.321-324
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    • 2001
  • Groundwater samples were collected at 6 sites in Seoul area. DNA extraction from the sample was performed by the boiling method. Samples were boiled with guanidinium thyocyanate and phenol-chloroform. One set of primer was designed for amplification of 16S rDNA. For detection of denitrifying bacteria in groundwater sample, the author used primer sets consensus regions in gene sequences encoding the two forms of nitrite reductase (NIR), a key enzyme in the denitrification pathway. Two sets of PCR primer were designed to amplify $cd_1$-and Cu-nir. We confirmed the existence of denitrifying bacteria in 3 sites using $cd_1$-nir primer and in 4 sites using Cu-nir primer.

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REWETTING EFFECT OF WATER-BASED PRIMER ON THE AIR-DRIED DENTIN (공기건조된 상아질에 대한 수분함유 primer의 재습윤효과)

  • Kim, Ki-Young;Park, Jeong-Kil;Hur, Bock
    • Restorative Dentistry and Endodontics
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    • v.28 no.6
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    • pp.498-503
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    • 2003
  • The purpose of this study was to evaluate the rewetting effect of water-based primer on the air-dried dentin. In this in vitro study, freshly extracted non-caries human molars and three-step adhesive system(SBMP) were used. Freshly extracted non-caries human molars and three-step adhesive system(SBMP) were used. Flat occlusal dentin surface were prepared using low-speed diamond saw, Prepared teeth were randomly divided into three groups. Group 1.(W): etched(35% phosphoric acid for 15s) and blot-dried, Group 2.(5D): 5s air-dried, Group 3.(30D): 30s ail-dried, To obtain color contrast in CLSM observation, primer was mixed with rhodamine B and bonding resin was mixed with fluorescein. Microscopic sample of each group were obtained after longitudinal section. Morphological investigation of resin-dentin interface and thickness of hybrid layer measurement using CLSM were done. Microtensile bond strength for each specimen was measured. Specimen were observed under microscope to examine the failure patterns of interface between resin and dentin. The results of this study were as follows: 1. The results(mean) of Thickness of hybrid layer were W:19.67, 5D:20.9, 30D:10$\mu\textrm{m}$. Only 30D had statistically significant differences to Wand 5D(P<0.05). 2. The results(mean) of Microtensile bond strength were W:16.02, 5D:14.69, 30D:11.14MPa. Only 30D had statistically significant differences to Wand 5D(P<0.05). 3. There were positive correlation between Thickness of hybrid layer and microtensile bond strength(P<0.05).