• Journal of Animal Science and Technology
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• 제46권6호
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• pp.897-902
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• 2004

이전에 연구된 한우육과 Holstein 및 Black angus육 감별법의 신속성, 편리성, 경제성 등의 단점을 보완하기 위해 소의 MC1R gene의 SNP를 이용한 새로운 감별법을 시도하였다. 본 연구에서는 소의 MC1R gene 중 594번째 염기인 Guanine이 한우에서는 결실된 점을 이용하여 한우의 sequence를 바탕으로 3 쪽에 2mers의 tail을 달아 한우에게는 상보적이나 다른 종에서는 상보적이지 않은 3 -tailed primers를 제작하였다. 이 primer들을 이용해서 한우에서만 MC1R 중 571번째 염기서열부터 919번째 염기서열까지의 343bp의 단편이 증폭되도록 하였다. 그 결과, Holstein, Black angus에서는 모두 band가 관찰되지 않았으나 한우에서는 343bp의 band가 확인되었다. 따라서 본 연구에서 사용한 3 -tailed primer를 이용하면 보다 정확하고 재현성 있으며 신속하고 편리하며 경제적인 한우육의 감별이 될 것으로 판단된다.

• The Journal of Advanced Prosthodontics
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• 제5권2호
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• pp.198-203
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• 2013

PURPOSE. The purpose of this study was to investigate the effect of resin cements and primer on the retentive force of zirconia copings bonded to zirconia abutments with insufficient retention. MATERIALS AND METHODS. Zirconia blocks (Lava, 3M ESPE, St. Paul, MN, USA) were obtained and forty sets of zirconia abutments and copings were fabricated using CAD/CAM technology. They were grouped into 4 categories as follows, depending on the types of resin cements used, and whether the primer is applied or not:Panavia F2.0 (P), Panavia F2.0 using Primer (PRIME Plus, Bisco Inc, Schaumburg, IL, USA) (PZ), Superbond C&B (S), and Superbond C&B using Primer (SZ). For each of the groups, the cementation was conducted. The specimens were kept in sterilized water ($37^{\circ}C$) for 24 hours. Retentive forces were tested and measured, and a statistical analysis was carried out. The nature of failure was recorded. RESULTS. The means and standard deviations of retentive force in Newton for each group were $265.15{\pm}35.04$ N (P), $318.21{\pm}22.24$ N (PZ), $445.13{\pm}78.54$ N (S) and $508.21{\pm}79.48$ N (SZ). Superbond C&B groups (S & SZ) showed significantly higher retentive force than Panavia F2.0 groups (P & PZ). In Panavia F2.0 groups, the use of primer was found to contribute to the increase of retentive force. On the other hand, in Superbond C&B groups, the use of primer did not influence the retention forces. Adhesive failure was observed in all groups. CONCLUSION. This study suggests that cementation of the zirconia abutments and zirconia copings with Superbond C&B have a higher retentive force than Panavia F2.0. When using Panavia F2.0, the use of primer increases the retentive force.

• Parasites, Hosts and Diseases
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• 제47권3호
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• pp.293-297
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• 2009

Improved methods for detection of Cryptosporidium oocysts in environmental and clinical samples are urgently needed to improve detection of cryptosporidiosis. We compared the sensitivity of 7 PCR primer sets for detection of Cryptosporidium parvum. Each target gene was amplified by PCR or nested PCR with serially diluted DNA extracted from purified C. parvum oocysts. The target genes included Cryptosporidium oocyst wall protein (COWP), small subunit ribosomal RNA (SSU rRNA), and random amplified polymorphic DNA. The detection limit of the PCR method ranged from $10^3$ to $10^4$ oocysts, and the nested PCR method was able to detect $10^0$ to $10^2$ oocysts. A second-round amplification of target genes showed that the nested primer set specific for the COWP gene proved to be the most sensitive one compared to the other primer sets tested in this study and would therefore be useful for the detection of C. parvum.

• 생명과학회지
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• 제17권1호
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• pp.162-165
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• 2007

Brown Ring Disease (BRD) is a bacterial disease caused by Vibrio tapetis which affects cultured clam Ruditapes philippinarum and causes heavy economic losses on Atlantic coasts of france, Spain and England. In this study, to evaluate the effective detection of the pathogen, specific primer set based on 16S ribosomal RNA (rRNA) sequences designed for rapid detection of V. tapetis. Polymerase chain reaction (PCR) with this primer set produced the specific band for each V. tapetis. The length of PCR product using designed primer set of Vbts-F and Vbts-R was about 400 bp. Therefore, these primers will be provided with a basic tool for rapid detection of V. tapetis in the various cases such as examination of imported aquatic products, diagnosis of aquatic organisms, and etc.

• 한국어병학회지
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• 제16권1호
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• pp.51-59
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• 2003

Streptococcal infection is one of the most serious disease of cultured olive flounder, Paralychthys olivaceus in Korea and caused by more than one species. However, there has been considerable confusions about the taxonomic position of the fish pathogenic streptococci. In this study, We performed the randomly amplified polymorphic DNA(RAPD) pattern analysis to evaluate the possible classification in 8 streptococci isolated from diseased olive flounder and reference strains based on their DNA structure. RAPD PCR with DNA solution prepared by simple boiling and 10-mer random primer was appeared to be a good tool for discrimination of different streptococcal strains. Phenotypical characters by simple biological test and API 20 Strep corresponded well to the specific profiles of RAPD in streptococcal isolates of this study. Therefore, the RAPD profile was considered as one of differential characters to discriminate the streptococcal isolates from diseased olive flounder.

• Journal of Plant Biotechnology
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• 제31권1호
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• pp.1-6
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• 2004

한약재로 사용되는 방풍류는 절단되어 유통되므로 외부 형태적인 특징만으로 구분하기가 어려워 방풍류로 사용되는 방풍, 식방풍, 석방풍, 갯방풍 등 4종에 대해 PCR에 기초한 RAPD 마커를 이용하여 SCAR 마커를 개발하고자 하였다. RAPD 분석결과 밴드의 패턴은 다양하게 나타났으며 다형성의 밴드 수는 총 215개로 전체 밴드수의 98%였다. RAPD 분석에서 각 방풍류를 구별 할 수 있는 특이적인 밴드를 나타내는 primer는 방풍에서 4개의 primer, 식방풍은 6개의 primer, 석방풍은 4개의 primer, 갯방풍은 6개의 primer를 선발하였고, 그 중 특히 primer 425는 4종의 방풍류의 감별에 유용하였고, 이를 이용하여 SCAR마커로 전환하는데 이용하였다. 특이적인 단편을 클로닝하여 염기서열 분석으로 특이 primer를 제작하고 제작된 primer로 방풍류 시료 16개에 적용하였을 때, 국내의 야생에서 주로 자생하는 석방풍은 215 bp, 그리고 국내에서 가장 많이 재배 또는 생산되는 갯방풍은 177 bp와 300 bp에서 뚜렷하게 나타났다. 따라서 갯방풍과 석방풍의 감별 가능성을 제시할 수 있으며 개발된 SCAR 마커를 이용하여 시중에 유통되고 있는 방풍류 건조약재의 감별에 유용한 마커로 활용될 수 있을 것이다.

• Parasites, Hosts and Diseases
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• 제49권3호
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• pp.281-284
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• 2011

Amebiasis is a protozoan disease caused by Entamoeba histolytica and a potential health threat in areas where sanitation and hygiene are inappropriate. Highly sensitive PCR methods for detection of E. histolytica in clinical and environmental samples are extremely useful to control amebiasis and to promote public health. The present study compared several primer sets for small subunit (SSU) rDNA and histone genes of E. histolytica cysts. A 246 bp of the SSU rDNA gene of pure cysts contained in phosphate-buffered saline (PBS) and in stool samples was successfully amplified by nested PCR, using the 1,147-246 bp primer set, of the primary PCR products which were pre-amplified using the 1,147 bp primer as the template. The detection limit of the nested PCR using the 1,147-246 primer set was 10 cysts in both groups (PBS and stool samples). The PCR to detect histone gene showed negative results. We propose that the nested PCR technique to detect SSU rDNA can be used as a highly sensitive genetic method to detect E. histolytica cysts in stool samples.

• 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.

• 대한한의학원전학회지
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• 제33권1호
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• pp.17-28
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• 2020

Objectives : Classical texts such as 『Donguibogam』 and 『Yixuerumen』 have previously been used as primers to students of Korean Medicine. However, their massiveness in volume and comprehensiveness in contents make it unfit for students whose school curriculum lacked classical chinese. This paper suggests another introductory text that would be more practical in the current situation. Methods :Based on the translation of the main text and annotations, the clinical meanings of the contents were studied. Afterwards its practical application as a primer was considered. Results : The text focuses on the medically important issues in simple and accessible form, making it an important text for beginners to establish the foundation in medicine. Conclusions : Beginners will be able to establish a standard for basic medical knowledge through this text and also apply its contents to diseases that are relatively easy to treat.

• The Plant Pathology Journal
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• 제32권1호
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• pp.53-57
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• 2016

Major diseases in grafted cacti have been reported and Fusarium oxysporum, Bipolaris cactivora, Phytophthora spp. and Collectotrichum spp. are known as causal pathogens. These pathogens can lead to plant death after infection. Therefore, some European countries have quarantined imported cacti that are infected with specific fungal pathogens. Consequently, we developed PCR detection methods to identify four quarantined fungal pathogens and reduce export rejection rates of Korean grafted cacti. The pathogen specific primer sets F.oF-F.oR, B.CF-B.CR, P.nF-P.nR, and P.cF-P.CR were tested for F. oxysporum, B.cactivora, P. nicotinae, and P. cactorum, respectively. The F.oF-F.oR primer set was designed from the Fusarium ITS region; the B.CF-B.CR and P.nF-P.nR primers respectively from Bipolaris and Phytophthora ITS1; and the P.cF-P.CR primer set from the Ypt1protein gene region. The quarantine fungal pathogen primer pairs were amplified to the specific number of base pairs in each of the following fungal pathogens: 210-bp (F. oxysporum), 510-bp (B. cactivora), 313-bp (P. nicotinae), and 447-bp (P. cactorum). The detection limit for the mono- and multiplex PCR primer sets was 0.1 ng of template DNA under in vitro conditions. Therefore, each primer set successfully diagnosed contamination of quarantine pathogens in export grafted cacti. Consequently, our methodology is a viable tool to screen contamination of the fungal pathogen in exported grafted cacti.