• Title/Summary/Keyword: Water based 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.

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).

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.

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

  • Jeong, Gwon-Young;Youn, Da Ae;Jang, Seok-Joon;Kil, Bae-Su;Yun, Hyun-Do
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.21 no.4
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    • pp.39-46
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    • 2017
  • The experimental research was conducted to evaluate the adhesive performance of surface finishing material with primer based on silane(primer). For this purpose, concrete specimens with compressive strength of 18, 30, 50 MPa were made and cured in water condition ($20{\pm}2^{\circ}C$) for 28 days. A primer was applied on the age of 28 days and evaluated according to based on the curing age of the surface finishing material. Moreover, the mortar specimen also made and tested as per KS F 4937 for compared with concrete-based test results. Test results indicated that the adhesive strength of specimens with primer exhibit similar than that of specimens without primer. Also, the adhesive performance improved with increasing in curing age and compressive strength. The correlation between compressive and adhesive strength of mortar and concrete specimens showed similar trend. It was noted that there is no significant effects of primer on adhesive performance of surface finishing material, thus use of primer has superior potential for solving durability problem of concrete slab surface.

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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    • v.22 no.8
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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
    • Biomedical Science Letters
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    • v.24 no.3
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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.

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

  • Kim, Young-Ung;You, Kwang-Ho;Cho, Nam-Hyun
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.36 no.4
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    • pp.695-704
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    • 2016
  • Rubber deposited during aircraft landing is known as the main cause of reducing surface friction force on wet surfaces. Thus, rubber deposits are removed at regular intervals for sae airplane landing. The high-pressure waterblast method, widely used for the removal of rubber deposits, is regarded as the main cause for the loss of surface material because in this method, water hits the surface directly at a high pressure. In this study, a rubber removal primer is developed to reduce surface damage by lowering the pressure of waterblast relatively during the removal of rubber deposits such that the deposits are removed efficiently even with a lower water pressure. To achieve this, basic materials appropriate for the primer were selected and their performance, penetration rate, and site applicability were evaluated. Based on the evaluations, the proportion of additive required for improving the performance of the basic materials was first determined. Then, the optimum mix ratio was derived through the evaluation of the effect on pavements, and the development of the rubber removal primer was completed.

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
    • Korean Journal of Agricultural Science
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    • v.47 no.3
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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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    • v.23 no.4
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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.