• 제목/요약/키워드: specific detection

검색결과 2,437건 처리시간 0.031초

Development of Nested PCR Primer Set for the Specific and Highly Sensitive Detection of Human Parvovirus B19

  • Cho, Kyu-Bong
    • 대한의생명과학회지
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    • 제24권4호
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    • pp.390-397
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    • 2018
  • For the specific detection of human Parvovirus B19 (HuPaV-B19), we designed ten specific PCR primers from 3,800~4,500 nucleotides of HuPaV-B19 complete genome (NC_000883.2). Seventeen candidate PCR primer sets for specific detecting HuPaV-B19 were constructed. In specific reaction of HuPaV-B19, seventeen PCR primer sets showed specific band, however five PCR primer sets were selected basis of band intensity, amplicon size and location. In non-specific reaction with seven reference viruses, four PCR primer sets showed non-specific band, however one PCR primer set is not. Detection sensitivity of final selective PCR primer set was $100fg/{\mu}L$ for 112 minute, and PCR amplicon is 539 base pairs (bp). In addition, nested PCR primer set was developed, for detection HuPaV-B19 from a low concentration of contaminated samples. Selection of nested PCR primer set was basis of sensitivity and groundwater sample tests. Detection sensitivity of final selective PCR and nested PCR primer sets for the detection of HuPaV-B19 were $100fg/{\mu}L$ and $100ag/{\mu}L$ basis of HuPaV-B19 plasmid, it was able to rapid and highly sensitive detection of HuPaV-B19 than previous reports. We expect developed PCR primer set in this study will used for detection of HuPaV-B19 in various samples.

봉변에서 특이 유전자 검출법에 의한 봉군 내 꿀벌가시응애류 (Tropilaelaps)의 정량적 검출 (Quantitative Detection of Tropilaelaps in Hive by Specific Gene Detection from Hive Debris)

  • 김병희;김소민;김문정;김정민;;김선미;윤병수
    • 한국양봉학회지
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    • 제34권1호
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    • pp.27-37
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    • 2019
  • Rapid detection of Tropilaelaps, an external parasite of honeybees that lead to malformation of honeybee or colony collapse disorder, is becoming important. But it is very difficult to find with the naked eye of Tropilaelaps. In this study, we have developed a method to detect the specific gene of Tropilaelaps from the hive debris and to know the number of Tropilaelaps in the hive through Tropilaelaps-specific quantitative detection. Tropilaelaps-specific gene amplified in DNA extracted from hive debris by consecutive PCR (1st detection, 2nd nested PCR). It could detect 101 molecules level of Tropilaelaps-specific gene and confirm the amplification of the Tropilaelaps-specific gene. It was possible to accurately quantify the number of Tropilaelaps from the hive debris sample, which is difficult to discriminate the presence of Tropilaelaps visually, through Tropilaelaps-specific detection. Under the microscope, Tropilaelaps was collected and quantitative detection of Tropilaelaps-specific genes was performed. It was possible to quantify the number of Tropilaelaps present in the hive through the molecules of the quantified Tropilaelaps-specific genes. We suggest that hive debris can represent as a micro-environment to hive and show that it can be a simpler and more accurate sample than using a parasitic host honeybee. We expect that hive debris should facilitate the monitoring of Tropilaelaps in hive.

Robust Face Detection Based on Knowledge-Directed Specification of Bottom-Up Saliency

  • Lee, Yu-Bu;Lee, Suk-Han
    • ETRI Journal
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    • 제33권4호
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    • pp.600-610
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    • 2011
  • This paper presents a novel approach to face detection by localizing faces as the goal-specific saliencies in a scene, using the framework of selective visual attention of a human with a particular goal in mind. The proposed approach aims at achieving human-like robustness as well as efficiency in face detection under large scene variations. The key is to establish how the specific knowledge relevant to the goal interacts with the bottom-up process of external visual stimuli for saliency detection. We propose a direct incorporation of the goal-related knowledge into the specification and/or modification of the internal process of a general bottom-up saliency detection framework. More specifically, prior knowledge of the human face, such as its size, skin color, and shape, is directly set to the window size and color signature for computing the center of difference, as well as to modify the importance weight, as a means of transforming into a goal-specific saliency detection. The experimental evaluation shows that the proposed method reaches a detection rate of 93.4% with a false positive rate of 7.1%, indicating the robustness against a wide variation of scale and rotation.

Specific Detection of Enteropathogen Campylobacter jejuni in Food Using a Polymerase Chain Reaction

  • Shin, Soon-Young;Park, Jong-Hyun;Kim, Wang-June
    • Journal of Microbiology and Biotechnology
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    • 제9권2호
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    • pp.184-190
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    • 1999
  • The use of the polymerase chain reaction (PCR) method was described using two sets of primers based on the ceuN gene (JEJ 1 and JEJ 2) which encodes a protein involved in siderophore transport and 16S rRNA gene (pA and pB) for the sensitive and specific detection of enteropathogen Campylobacter jejuni. Six oligonucleotides were utilized in an amplification experiment and PCR products of predicted sizes were generated from whole cells and boiled cell lysates at the same intensity. Two sets of the primer pairs, JEJ and pAB, were specific enough for all C. jejuni strains tested for the direct use of whole cells without DNA extraction or lysis steps. In the PCR using the pAB primer pair, the detection limit, as determined by the ethidium bromide staining of the amplification products on agarose gels, was at the level of $10^1$ bacteria cells or less in both the pure culture and artificially inoculated milk and chicken enrichment samples, whereas the detection limit with the JEJ primer pair was relatively low, i.e. $10^3$ cells or more in the same PCR samples. The PCR method using either a primer JEJ or pAB was both repeatable and specific for the detection of C. jejuni in food. This method is simply completed within 4 h.

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초분광 영상 특징선택과 밴드비 기법을 이용한 유사색상의 특이재질 검출기법 (Specific Material Detection with Similar Colors using Feature Selection and Band Ratio in Hyperspectral Image)

  • 심민섭;김성호
    • 제어로봇시스템학회논문지
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    • 제19권12호
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    • pp.1081-1088
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    • 2013
  • Hyperspectral cameras acquire reflectance values at many different wavelength bands. Dimensions tend to increase because spectral information is stored in each pixel. Several attempts have been made to reduce dimensional problems such as the feature selection using Adaboost and dimension reduction using the Simulated Annealing technique. We propose a novel material detection method that consists of four steps: feature band selection, feature extraction, SVM (Support Vector Machine) learning, and target and specific region detection. It is a combination of the band ratio method and Simulated Annealing algorithm based on detection rate. The experimental results validate the effectiveness of the proposed feature selection and band ratio method.

Development of strain-specific polymerase chain reaction primers to detect Fusobacterium hwasookii strains

  • Lim, Yun Kyong;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • 제46권4호
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    • pp.155-159
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    • 2021
  • This study aimed to develop strain-specific polymerase chain reaction (PCR) primers to detect Fusobacterium hwasookii KCOM 1249T, F. hwasookii KCOM 1253, F. hwasookii KCOM 1256, F. hwasookii KCOM 1258, and F. hwasookii KCOM 1268 on the basis of nucleotide sequences of a gene specific to each strain. The unique genes for each F. hwasookii strain were determined on the basis of their genome sequences using Roary. The strain-specific PCR primers based on each strain-specific gene were designed using PrimerSelect. The specificity of each PCR primer was determined using the genomic DNA of the 5 F. hwasookii strains and 25 strains of oral bacterial species. The detection limit and sensitivity of each strain-specific PCR primer pair were determined using the genomic DNA of each target strain. The results showed that the strain-specific PCR primers correspond to F. hwasookii KCOM 1249T, F. hwasookii KCOM 1253, F. hwasookii KCOM 1258, F. hwasookii KCOM 1256/F. nucleatum subsp. polymorphum KCOM 1260, or F. hwasookii KCOM 1268/Fusobacterium sp. oral taxon 203 were developed. The detection limits of these strain-specific PCR primers ranged from 0.2 to 2 ng of genomic DNA for each target strain. The results suggest that these strain-specific PCR primers are valuable in quality control for detecting specific F. hwasookii strains.

Development of Species-Specific PCR Primers for the Detection of Streptococcus sobrinus

  • Kim, Sang-Gon;Yoo, So-Young;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • 제35권1호
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    • pp.21-25
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    • 2010
  • This study was undertaken to develop species-specific forward and universal reverse PCR primers for the detection of Streptococcus sobrinus. These primers target the variable regions of the 16S ribosomal RNA coding gene (rDNA) and their specificity was tested against 10 strains of S. sobrinus strains and 20 different species of oral bacteria using serial dilutions of the purified genomic DNA of S. sobrinus ATCC $33478^T$. Our data show that species-specific amplicons were obtained from all the S. sobrinus strains tested but not from other species. Both direct and nested PCR could detect as little as 400 pg and 4 fg of genomic DNA from S. sobrinus ATCC $33478^T$, respectively. This result suggests that these PCR primers are highly specific and sensitive and applicable to the detection of S. sobrinus.

맥주오염미생물의 동정과 specific PCR primer의한 신속한 검출 방법 (Characterization of beer-spoilage microorganism and its rapid detection by specific PCR primer)

  • 이택인;최신건
    • 산업기술연구
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    • 제28권A호
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    • pp.141-147
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    • 2008
  • Several contaminated bacteria such as Lactobacillus brevis and Pediococcus damnosus in beer production cause beer spoilage by producing off flavours and turbidity. Detection of these organisms is complicated by the strict anaerobic conditions and lengthy incubation times required for their cultivation, consequently there is a need for more rapid detection methods. Recently, two contaminated strains were isolated from vessel of beer production and identified as Lactobacillus species by API kit identificaton as well as 16S-23S ITS sequencing analyses. Two isolated strains were named as Lactobacillus sp. HLA1 and Lactobacillus HLB2, respectively. A polymerase chain reaction (PCR) method was developed for the rapid and specific detection of Lactobacillus sp.. Two sets of primer pairs (HLA1-F/HLA1-R and HLB2-F/HLB2-R) were designed for the amplification of a 1576 base pair (bp) fragment of the HLA1 16S-23S rRNA gene and 1888 bp fragement of the HLB2 16S-23S rRNA. Amplified PCR products were highly specific to detect corresponding bacteria when other contaminated strains were used as PCR templates. However, detection of both strains were limited when $100{\mu}{\ell}$ of cultured samples were mixed with $100m{\ell}$ of beer sample in arbitrary manner. The sensitivity of the assay still needs to be improved for direct detection of the small amounts of bacteria present in beer.

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중합효소연쇄반응(PCR)을 이용한 고양이 혈액내에서의 Toxoplasma gondii 검출에 관한 연구 (Polymerase chain reaction for the detection of Toxoplasma gondii in the blood of cats)

  • 서명득;주보현
    • 대한수의학회지
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    • 제39권6호
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    • pp.1151-1160
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    • 1999
  • This study was conducted to detect the toxoplasma-specific DNA in peripheral blood collected from cats experimentally infected with Toxoplasma gondii (RH strain) and from domiciled cats by B1 gene-base polymerise chain reaction(PCR). The sensitivity of oligonucleotide primer, T-1 & T-2, designed from toxoplasma B1 gene amplification method was compared with parasite detection by mouse inoculation(MI). And also, latex agglutination test(LAT) and indirect fluorescent antibody test(IFAT) were conducted to detect the fluctuation of serum antibodies compared with the detection of toxoplasma by PCR and MI. Toxoplasma B1 gene PCR was shown consistently high sensitivity and the results obtained by PCR agreed completely with those from MI. All blood samples collected before infection with T gondii gave negative results by PCR and MI. Also, toxoplasma Bl gene PCR was not cross reaction with Neospora caninum DNA and normal cat leucocyte as controls. The toxoplasma-specific DNA was detected by PCR in blood of 5 cats experimentally infected with T gondii 6 days after infection and the detection of this specific-DNA was long lasted in blood for 64 days after infection. The detection of toxoplasma-specific DNA by PCR could be identified as few as 10 tachyzoites and the isolation of T gondii by MI could be isolated as few as 1 tachyzoite from tenfold serial dilution of T gondii with normal cat blood, respectively. In healthy domiciled cats, the toxoplasma-specific DNA and the parasite were detected and isolated in blood from 3 of 56(5.3%) cats by both PCR and MI, respectively. In the results of antibody test from the total 56 heads of healthy domiciled cats, the positive rates are 15(26.7%) by LAT and 19(33.9%) by IFAT. These results suggest that PCR detection of toxoplasma can be applied as a sensitive and specific diagnostic and research tool.

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수동소나시스템에서 탐지효과도 분석 (Measure of Effectiveness Analysis of Passive SONAR System for Detection)

  • 조정홍;김재수
    • 한국군사과학기술학회지
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    • 제15권3호
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    • pp.272-287
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    • 2012
  • The optimal use of sonar systems for detection is a practical problem in a given ocean environment. In order to quantify the mission achievability in general, measure of effectiveness(MOE) is defined for specific missions. In this paper, using the specific MOE for detection, which is represented as cumulative detection probability(CDP), an integrated software package named as Optimal Acoustic Search Path Planning(OASPP) is developed. For a given ocean environment and sonar systems, the discrete observations for detection probability(PD) are used to calculate CDP incorporating sonar and environmental parameters. Also, counter-detection probability is considered for vulnerability analysis for a given scenario. Through modeling and simulation for a simple case for which an intuitive solution is known, the developed code is verified.