• 생명과학회지
• /
• 제17권1호
• /
• pp.162-165
• /
• 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.

• 한국식품영양과학회지
• /
• 제28권5호
• /
• pp.1082-1086
• /
• 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.

• Journal of Biosystems Engineering
• /
• 제39권2호
• /
• pp.111-114
• /
• 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.

• Fisheries and Aquatic Sciences
• /
• 제16권4호
• /
• pp.273-277
• /
• 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.

• 한국정보처리학회논문지
• /
• 제5권3호
• /
• pp.715-719
• /
• 1998

본 논문에서는 TV에 있어서 위상 정보를 이용하여 특정색을 조정할 경우 색신호의 천이 시간에서 발생하는 검출 오류들과 색신호의 안정화를 위한 색향상 시스템을 제안하였다. 제안한 색향상 시스템은 시간차 보정 방법을 적용하여 색신호와 색부 반송 신호의 전송 과정에서 발생하는 시간차를 보정하여 색신호 검출 오류를 줄였다. 또한 변화하는 색신호의 위상을 검출함에 있어서 실제적으로 존재하는 천이 시간에 따른 검출 오류를 개선하고자 수평 동기 시간내에서 검출 전압값의 평균 차성분을 구하여 이웃한 색신호의 최소 구별 전압값보다 큰값을 가질 때 설정 기준색을 조정하도록 하는 안정화 방법을 제안하였다. 제안한 개선된 색향상 시스템은 기타의 색신호로부터 보정 범위가 중복되지 않고 특정색의 검출 및 조정이 개선되었다.

• The Plant Pathology Journal
• /
• 제21권4호
• /
• pp.343-348
• /
• 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.

• Journal of Microbiology and Biotechnology
• /
• 제18권9호
• /
• pp.1492-1495
• /
• 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.

• Journal of Microbiology and Biotechnology
• /
• 제16권11호
• /
• pp.1778-1783
• /
• 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.

• The Plant Pathology Journal
• /
• 제16권5호
• /
• pp.252-256
• /
• 2000

The 16S-23S rRNA intergenic spacer regions (ISRs) were sequenced and analyzed to design specific primer for identification of Pectobacterium chrysanthemi. Two types ISRs, large and small ISRs, were identified from three strains (ATCC 11663, KACC 10163 and KACC 10165) of P. chrysanthemi and Pectobacterium carotovorum subsp. carotovorum ATCC 15713.Large ISRs contained transfer RNA-Ile(tRNA$^{Ile}$)and tRNA$^{Ala}$, and small ISRs contained tRNA$^{Glu}$. Size of the small ISRs of P. chrysanthemi ranged on 354-356 bp, while it was 451 bp in small ISR of P. carotovorum subsp. carotovorum ATCC 15713. From hypervariable region of small ISRs, species-specific primer for P. chrysanthemi with 20 bp length (CHPG) was designed from hypervariable region of small ISRs, which was used as forward promer to detect P. chrysanthemi strains with R23-1R produced PCR product of about 260bp size (CHSF) only from P. chrysanthemi strains, not from other Pectobacterium spp. and Erwinia spp. Direct PCR from bacterial cell without extracting DNA successfully amplified a specific fragment, CHSF, from P. chrysanthemi ATCC 11663. The limit of PCR detection was 1${\pm}10^2$ cfu/ml.

• Food Science and Biotechnology
• /
• 제18권5호
• /
• pp.1118-1123
• /
• 2009

Event-specific real-time polymerase chain reaction (PCR) detection method for genetically modified (GM) maize MIR604 was developed based on integration junction sequences between the host plant genome and the integrated transgene. In this study, 2 primer pairs and probes were designed for specific amplification of 100 and 111 bp DNA fragments from the zSSIIb gene (the maize endogenous reference gene) and MIR604. The quantitative method was validated using 3 certified reference materials (CRMs) with levels of 0.1, 1, and 10% MIR604. The method was also assayed with 14 different plants and other GM maize. No amplification signal was observed in real-time PCR assays with any of the species tested other than MIR604 maize. As a result, the bias from the true value and the relative deviation for MIR604 was within the range from 0 to 9%. Precision, expressed as relative standard deviation (RSD), varied from 2.7 to 10% for MIR604. Limits of detections (LODs) of qualitative and quantitative methods were all 0.1%. These results indicated that the event-specific quantitative PCR detection system for MIR604 is accurate and useful.