• 제목/요약/키워드: Water based primer

검색결과 49건 처리시간 0.023초

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

  • 김광일;진지웅;정현도
    • 한국어병학회지
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    • 제24권3호
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    • pp.255-268
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    • 2011
  • 본 연구에서는 RT-nested PCR을 수행하여 부산 도심의 하천 중 동천에 존재하는 노로바이러스를 검출하고자 하였다. 기존에 보고되어진 노로바이러스의 capsid protein의 염기서열을 비교하여 노로바이러스 genogroup I,II (GI,II) 를 검출하기 위한 새로운 degenerated primer sets (PNK, KGIF/KGIR and KG2F/KG2R) 를 제작하였으며, 채수한 동천 시료를 초고속원심분리기를 통해 농축 후 물 속에 존재하는 노로바이러스의 검출을 시도하였다. 노로바이러스를 검출하기 위해 PCR primer를 비교한 결과 본 연구에서 제작한 capsid protein gene을 target으로 하는 primer set가 기존에 보고되어 있는 primer set보다 동일 시료에 대한 검출빈도가 우수하였다. 동천에 존재하는 노로바이러스의 오염 수준은 GI과 GII가 각각 76.47% (13/17), 70.59% (12/17) 로 나타났다. 그러나 기존에 알려진 primer와 본 연구에서 제작한 primer를 사용하였을 때 검출된 양성비율이 차이가 나지 않았다. 검출된 노로바이러스를 염기서열 비교를 통한 계통 발생학적 분석 결과, 동천에서 검출된 GI의 경우 1/2/4/5/9/10의 genotype이 GII의 경우 3/4/5/11/13의 genotype으로 분류되었다. 그리고 본 연구에서 검출된 major type 중 GII/4의 경우, 최근 아시아 각국에서 많은 문제를 일으키고 있는 major genotype으로 알려져 노로바이러스에 대한 위험성을 제고하게 하였다. 또한, 이러한 결과는 국내의 강, 호수, 하천 등이 비슷한 노로바이러스의 GI,II의 genotype으로 오염되어 있음을 암시하며 수계환경 중 미생물의 질을 개선하기 위한 지속적인 노로바이러스의 모니터링이 요구된다.

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

  • 김기영;박정길;허복
    • Restorative Dentistry and Endodontics
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    • 제28권6호
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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).

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
    • 농업과학연구
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    • 제47권1호
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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.

실란계 프라이머를 활용한 바닥 마감재 부착성능 평가 (Evaluation of Adhesive Performance of Surface Finishing Material with Primer Based on Silane)

  • 정권영;윤다애;장석준;길배수;윤현도
    • 한국구조물진단유지관리공학회 논문집
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    • 제21권4호
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    • pp.39-46
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    • 2017
  • 본 연구에서는 프라이머 도포 여 부 및 압축강도수준 그리고 바닥 마감재 시공재령에 따른 주차장용 바닥 마감재 부착성능을 평가하였다. 또한 현행 KS 기준에서는 바닥 마감재 부착성능을 모르타르 공시체를 활용하여 평가하도록 제안하고 있지만, 실제 현장에서 바닥 마감재는 콘크리트 상부면에 시공되기 때문에 바탕면 변화에 따른 부착성능을 추가변수로 계획하여 평가를 진행하였다. 모르타르는 현행기준에서 제안하고 있는 배합표에 준하여 제작하였으며, 콘크리트 설계기준강도는 18, 30, 50 MPa로 계획하였다. 프라이머는 모르타르 및 콘크리트 재령 28일 변수에 따라 바닥 마감재와 함께 도포되었으며, KS 기준에 준하여 바닥 마감재 재령 일에 따라 부착성능을 평가하였다. 변수에 따른 부착성능 평가결과 바닥 마감재 재령이 경과함에 따라 바닥 마감재 부착성능은 향상되는 것으로 나타났으며, 압축강도가 높아짐에 따라 바닥 마감재 부착성능 또한 향상되는 것으로 나타났다. 이는 파괴양상을 고려하였을 때 콘크리트 쪼갬 인장강도의 영향을 받은 것으로 판단된다. 프라이머 도포에 따른 부착성능은 유사한 것으로 나타났으며, 프라이머의 사용은 부착강도에 문제없이 콘크리트 표면의 내구성을 향상시킬 수 있을 것으로 판단된다. 또한 콘크리트 및 모르타르 바탕면의 부착강도 특성을 비교한 결과 바닥재의 부착강도는 구성재료 보다 압축강도에 큰 영향을 받는 것으로 나타났다. 따라서 현행 KS 기준의 모르타르 바탕면 실험체의 부착강도를 근거로 실제현장의 부착강도를 예측할 수 있을 것으로 판단된다.

A Novel Marker for the Species-Specific Detection and Quantitation of Shigella sonnei by Targeting a Methylase Gene

  • Cho, Min Seok;Ahn, Tae-Young;Joh, Kiseong;Kwon, Oh-Sang;Jheong, Won-Hwa;Park, Dong Suk
    • Journal of Microbiology and Biotechnology
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    • 제22권8호
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    • pp.1113-1117
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    • 2012
  • Shigella sonnei is a causal agent of fever, nausea, stomach cramps, vomiting, and diarrheal disease. The present study describes a quantitative polymerase chain reaction (qPCR) assay for the specific detection of S. sonnei using a primer pair based on the methylase gene for the amplification of a 325 bp DNA fragment. The qPCR primer set for the accurate diagnosis of Shigella sonnei was developed from publically available genome sequences. This quantitative PCR-based method will potentially simplify and facilitate the diagnosis of this pathogen and guide disease management.

Construction of Improved PCR Primer Set for the Detection of Human Enteric Adenovirus 41

  • Cho, Kyu-Bong
    • 대한의생명과학회지
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    • 제24권3호
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    • pp.230-238
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    • 2018
  • Human enteric Adenovirus-41 (HuEAdV-41) causes gastroenteritis, which detected by the polymerase chain reaction (PCR) base diagnostic system for clinical, food, environmental, fish and shellfish samples. We developed improved PCR and nested PCR primer set which had high specificity, sensitivity and reduced times. In this study, we compared seventeen conditions reported in the previous study that was using the PCR based HuEAdV-41 detection system, and non-enteric Adenovirus were detected in nine conditions. The most sensitive detection condition was up to 25 copies however it took 184 minutes of PCR reaction time. In this study, the PCR primer set developed had same level of sensitivity, it reduced the time of detection for clinical, food and seafood samples to 112 minutes. Developed nested PCR primer set needed 112 minutes but detected up to approximately 1 copy. In addition, developed PCR and nested PCR primer set was validated with twenty samples of underground water at random, of which ten samples showed specific band without non-specific reaction. We expect this study will be used to diagnose HuEAdV-41 from various samples.

활주로 고무 퇴적물 제거를 위한 포장 파손 저감형 사전처리제 개발 연구 (Development of the Rubber Removal Primer to Reduce Pavement Damage for Removal of Rubber Deposits in Runways)

  • 김영웅;유광호;조남현
    • 대한토목학회논문집
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    • 제36권4호
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    • pp.695-704
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    • 2016
  • 항공기 착륙 시 발생하는 고무퇴적물은 젖은 노면에서의 표면 마찰력을 감소시키는 주원인으로 안전한 항공기 착륙을 위해 주기적인 제거를 실시하고 있다. 제거작업에 주로 사용되는 고압살수 방법은 고압의 물로 직접 표면을 타격함에 따라 표면 재료 유실의 원인이 되고 있다. 본 연구에서는 고무 제거시 살수 압을 상대적으로 낮추어 표면 파손을 저감시키고, 저수압에도 효율적으로 고무퇴적물을 제거할 수 있는 사전 처리제 개발을 진행하였다. 이를 위해 사전 처리제에 적합한 기초재료를 선정하여 성능 평가, 침투율 평가, 현장 적용성 평가를 진행하였다. 이를 토대로 기초재료의 성능 개선에 필요한 첨가제의 비율을 1차적으로 선정하였고, 포장 영향성 평가를 통해 최적배합을 도출하여 고무제거 사전처리제 개발을 완료하였다.

Development of a diagnostic method for human enteric Adenovirus-41 with rapid, specific and high sensitivity using the loop-mediated isothermal amplification assay

  • Lee, Jin-Young;Rho, Jae Young
    • 농업과학연구
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    • 제47권3호
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    • pp.673-681
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    • 2020
  • Human enteric Adenovirus 41 (HueAdV-41) is a major waterborne virus that causes human gastroenteritis and is classified as a viral group I double-strand DNA virus, Adenoviridae. HueAdV-41 has been detected with the polymerase chain reaction (PCR) in various samples such as ground water. However, the PCR-based diagnostic method has problems such as reaction time, sensitivity, and specificity. Thus, the loop-mediated isothermal amplification (LAMP) assay has emerged as an excellent method for field applications. In this study, we developed a LAMP system that can rapidly detect HueAdV-41 with high specificity and sensitivity. HueAdV-41 specific LAMP primer sets were tested through a specific, non-specific selection and sensitivity test for three prepared LAMP primer sets, of which only one primer set and optimum reaction temperature were selected. The developed LAMP primer set condition was confirmed as 63℃, and the sensitivity was 1 copy. In addition, to confirm the system, a LAMP positive reaction was developed with the restriction enzyme Taq I (T/GCC). The developed method in this study was more specific, rapid (typically within 2 - 3 hours), and highly sensitive than that of the conventional PCR method. To evaluate and verify the developed LAMP assay, an artificial infection test was done with five cDNAs from groundwater samples, and the results were compared to those of the conventional PCR method. We expect the developed LAMP primer set will be used to diagnose HueAdV-41 from various samples.

A Novel Marker for the Species-Specific Detection and Quantitation of Vibrio cholerae by Targeting an Outer Membrane Lipoprotein lolB Gene

  • Cho, Min Seok;Ahn, Tae-Young;Joh, Kiseong;Paik, Soon-Young;Kwon, Oh-Sang;Jheong, Won-Hwa;Joung, Yochan;Park, Dong Suk
    • Journal of Microbiology and Biotechnology
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    • 제23권4호
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    • pp.555-559
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    • 2013
  • Vibrio cholerae O1 and O139 are the major serotypes associated with illness, and some V. cholera non-O1 and non-O139 isolates produce cholera toxin. The present study describes a quantitative polymerase chain reaction (qPCR) assay for the species-specific detection and quantitation of V. cholera using a primer pair based on an outer membrane lipoprotein lolB gene for the amplification of a 195 bp DNA fragment. The qPCR primer set for the accurate diagnosis of V. cholera was developed from publically available genome sequences. This quantitative PCR-based method will potentially simplify and facilitate the diagnosis of this pathogen and guide disease management.