• Title/Summary/Keyword: specific detection.

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Application of SYBR Green real-time PCR assay for the specific detection of Salmonella spp. (Salmonella spp. 특이적인 검출을 위한 SYBR Green real-time PCR 기법 적용)

  • Shin, Seung Won;Cha, Seung Bin;Lee, Won-Jung;Shin, Min-Kyoung;Jung, Myunghwan;Yoo, Anna;Jung, Byeng Yeal;Yoo, Han Sang
    • Korean Journal of Veterinary Research
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    • v.53 no.1
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    • pp.25-28
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    • 2013
  • The aim of this study was to applicate and evaluate a SYBR Green real-time PCR for the specific detection of Salmonella spp. Specificity of the PCR method was confirmed with 48 Salmonella spp. and 5 non-Salmonella strains using invA gene primer. The average threshold cycle ($C_T$) of Salmonella spp. was $11.83{\pm}0.78$ while non-Salmonella spp. was $30.86{\pm}1.19$. Correlation coefficients of standard curves constructed using $C_T$ versus copy number of Salmonella Enteritidis ATCC 13076 showed good linearity ($R^2=0.993$; slope = 3.563). Minimum level of detection with the method was > $10^2$ colony forming units (CFU)/mL. These results suggested that the SYBR Green real-time PCR might be applicable for the specific detection of Salmonella spp. isolates.

Development of Species-specific Primers for Rapid Detection of Phellinus linteus and P. baumii

  • Kim, Mun-Ok;Kim, Gi-Young;Nam, Byung-Hyouk;Jin, Cheng-Yun;Lee, Ki-Won;Park, Jae-Min;Lee, Sang-Joon;Lee, Jae-Dong
    • Mycobiology
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    • v.33 no.2
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    • pp.104-108
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    • 2005
  • Genus Phellinus taxonomically belongs to Aphyllophorales and some species of this genus have been used as a medicinal ingredients and Indian folk medicines. Especially, P. linteus and morphological-related species are well-known medicinal fungi that have various biological activities such as humoral and cell-mediated, anti-mutagenic, and anti-cancer activities. However, little is known about the rapid detection for complex Phellinus species. Therefore, this study was carried out to develop specific primers for the rapid detection of P. linteus and other related species. Designing the species-specific primers was done based on internal transcribed spacer sequence data. Each primer set detected specifically P. linteus (PL2/PL5R) and P. baumii (PB1/PB4R). These primer sets could be useful for the rapid detection of specific-species among unidentified Phellinus species. Moreover, restriction fragment length polymorphism analysis of the ITS region with HaeIII was also useful for clarifying the relationship between each 5 Phellinus species.

PCR Specific Primer for the Detection of Vibrio tapetis (Vibrio tapetis의 검출을 위한 PCR specific primer의 제작)

  • Kim, Yeong-Jin;Lee, Sun-Yi;Cho, Hyo-Jin;You, Sun-Nyung;Kim, Cheol-Min;Choi, Yong-Lark;Park, Byoung-Keun;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.17 no.1 s.81
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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.

Potential Detection of Irradiated Dried Agricultural Products by Viscosity Measurement (점도측정법을 이용한 방사선 조사 건조농산품의 검지 가능성)

  • 권중호;정형욱;정재영
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.28 no.5
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    • pp.1082-1086
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    • 1999
  • Dried vegetables, white ginseng and spices, which were exposed to gamma and electron beam irradiation, were used in a detection study by measuring their starch content and viscosity change. The samples tested showed different levels of starch content(15.64~60.86%), which was not directly proportional to the viscosity of the samples. The correlation coefficients between irradiation dose and viscosity change were lower in the samples, such as cabbage, carrot, clean vegetable(chunggyungchae), garlic, mushroom, green onion, and red pepper, while some higher coefficients were found in ginger(R2=0.9271), white ginseng (R2=0.6223) and onion (R2=0.7909). Thus, dried ginger and white ginseng were selected to be used for a detection of irradiated samples using specific parameters(threshold values). Specific parameter for the nonirradiated ginger and ginseng were 13.31 and 13.93, respectively. On the other hand, gamma and electron beam irradiated samples at 2.5 kGy, the lowest dose for a commercial purpose, showed decreased values, 11.92 and 11.15 in ginger, and moreover 4.40 and 5.10 in ginseng. It is expected that a proportional decrease in a specific parameter with the absorbed doses will be a potentially useful index for detecting whether starchy foods have been irradiated or not.

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The Importance of FACS Analysis in the Development of Aptamers Specific to Pathogens

  • Moon, Ji-Hea;Kim, Giyoung;Park, Saet Byeol;Lim, Jongguk;Mo, Changyeun
    • Journal of Biosystems Engineering
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    • v.39 no.2
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    • pp.111-114
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    • 2014
  • Purpose: This review aims to introduce aptamers and the methods of its development to improve the sensitivity and selectivity to target bacteria. In this review, we have highlighted current developments and directions in the pathogen detection based on aptamers. Background: Aptamers, the specific nucleic acid sequences, can bind to targets with high affinity and specificity. Some of researches on the use of aptamers for the detection of pathogen have been reported in recent years. Aptamers have more applicability than antibodies for the development of pathogen detection using biosensor; such as easy to synthesis and labeling, lack of immunogenicity, and a low cost of production. However, only few reports on the development and use of aptamers for the detection of pathogen have been published. Review: Aptamers specific to pathogen are obtained by whole-cell systematic evolution of ligands by exponential enrichment (SELEX) process. SELEX process is composed of screening random oligonucleotide bound with target cells, multiple separation and amplification of nucleic acids, final identification of the best sequences. For improving those affinity and selectivity to target bacteria, optimization of multiple separating process to remove unbounded oligonucleotides from aptamer candidates and sorting process by flow cytometry are required.

Species-Specific Duplex PCR for Detecting the Important Fish Pathogens Vibrio anguillarum and Edwardsiella tarda

  • Jo, Geon-A;Kwon, Sae-Bom;Kim, Na-Kyeong;Hossain, Muhammad Tofazzal;Kim, Yu-Ri;Kim, Eun-Young;Kong, In-Soo
    • Fisheries and Aquatic Sciences
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    • v.16 no.4
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    • pp.273-277
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    • 2013
  • Vibriosis caused by Vibrio anguillarum and edwardsiellosis caused by Edwardsiella tarda are septicemic diseases of many commercially important freshwater and marine fishes, and threaten the aquaculture industry in Korea. Early diagnosis and accurate identification of these two bacterial species could help to prevent these diseases and minimize the damage to cultured marine species. This study designed a duplex polymerase chain reaction (PCR) method for the simultaneous detection of two major fish pathogens: V. anguillarum and E. tarda. Each pair of oligonucleotide primers exclusively amplified the target groEL gene of the specific microorganism. Twenty-two Vibrio and ten non-Vibrio enteric species were used to check the specificity of the primers, which were found to be highly specific for the target species, even among closely related species. The detection limit was 400 pg for V. anguillarum and 4 ng for E. tarda when mixed purified DNA was used as the template. This assay showed high specificity and sensitivity in the simultaneous detection of V. anguillarum and E. tarda from artificially inoculated seawater and fish.

Color enhancement system Based on the improvement of Transition Time and color detdetion Stability (천이 시간 개선과 색검출 안정화 기반의 색 향상 시스템)

  • Lee, Eung-Joo
    • The Transactions of the Korea Information Processing Society
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    • v.5 no.3
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    • pp.715-719
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    • 1998
  • In this paper, we propose a color enhancement system which is based on the improvement of transition time problem and specific color detection stability. The proposcd system apply the time difference correction step to corrects time difference which is taken place at the transimission process between color signal and color subcarrier signal and to reduce detection errors. And also, we proposed stability method to improve transition time problem and detection efficiency as the control of reference color. The proposed system controls specific color when the mean difference value of detected voltages greater than the value of minimum discriminate voltages of two adjacent color signals. Thus, the color enhancement system improves detection efficiency and controls specific color from the color signal without overlapping of correction range.

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RT-PCR Detection of dsRNA Mycoviruses Infecting Pleurotus ostreatus and Agaricus blazei Murrill

  • Kim, Yu-Jeong;Park, Sang-Ho;Yie, Se-Won;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.21 no.4
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    • pp.343-348
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    • 2005
  • The partial nucleotide sequences of the genomic dsRNA mycoviruses infecting Pleurotus ostreatus (isolates ASI2596, ASI2597, and Bupyungbokhoe) and Agaricus blazei Murrill were determined and compared with those of the other dsRNA mycoviruses. Partial nucleotide sequences of the purified dsRNA from ASI2596 and ASI2597 revealed RNA-dependent RNA polymerase sequences that are closely related to Oyster mushroom isometric virus 2, while nucleotide sequences and the deduced amino acid sequence from dsRNA mycovirus infecting Agaricus blazei did not show any significant homology to the other dsRNA mycoviruses. Specific primers were designed for RT-PCR detection of these dsRNA viruses and were found to specifically detect each dsRNA virus. Northern blot analysis confirmed the homogeneity of RT-PCR products to each purified dsRNA. Altogether, our results suggest that these virus-specific primer sets can be employed for the specific detection of each dsRNA mycovirus in infected mushrooms.

Specific Detection of Xanthomonas oryzae pv. oryzicola in Infected Rice Plant by Use of PCR Assay Targeting a Membrane Fusion Protein Gene

  • Kang, Man-Jung;Shim, Jae-Kyung;Cho, Min-Seok;Seol, Young-Joo;Hahn, Jang-Ho;Hwang, Duk-Ju;Park, Dong-Suk
    • Journal of Microbiology and Biotechnology
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    • v.18 no.9
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    • pp.1492-1495
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    • 2008
  • Successful control of Xanthomonas oryzae pv. oryzicola, the causal agent of bacterial leaf streak, requires a specific and reliable diagnostic tool. A pathovar-specific PCR assay was developed for the rapid and accurate detection ofthe plant pathogenic bacterium Xanthomonas oryzae pv. oryzicola in diseased plant. Based on differences in a membrane fusion protein gene of Xanthomonas oryzae pv. oryzicola and other microorganisms, which was generated from NCBI (http://www.ncbi.nlm.nih.gov/) and CMR (http://cmr.tigr.org/) BLAST searches, one pair of pathovar-specific primers, XOCMF/XOCMR, was synthesized. Primers XOCMF and XOCMR from a membrane fusion protein gene were used to amplity a 488-bp DNA fragment. The PCR product was only produced from 4 isolates of Xanthomonas oryzae pv. oryzicola among 37 isolates of other pathovars and species of Xanthomonas, Pectobacterium, Pseudomonas, Burkholderia, Escherichia coli, and Fusarium oxysporum f.sp. dianthi. The results suggested that the assay detected the pathogen more rapidly and accurately than standard isolation methods.

Quantification of Genetically Modified Canola GT73 Using TaqMan Real-Time PCR

  • Kim, Jae-Hwan;Song, Hee-Sung;Kim, Dong-Hern;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1778-1783
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    • 2006
  • Event-specific PCR detection methods are the primary trend in genetically modified (GM) plant detection owing to their high specificity based on the flanking sequence of the exogenous integrant. Therefore, this study describes a real-time PCR system for event-specific GM canola GT73, consisting of a set of primers, TaqMan probe, and single target standard plasmid. For the specific detection of GT73 canola, the 3'-integration junction sequence between the host plant DNA and the integrated specific border was targeted. To validate the proposed method, test samples of 0, 1, 3, 5, and 10% GT73 canola were quantified. The method was also assayed with 15 different plants, and no amplification signal was observed in a real-time PCR assay with any of the species tested, other than GT73 canola.