• Title/Summary/Keyword: Species detection

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Highly Sensitive Fluorescent Probes for the Quantitative Determination of Singlet Oxygen (1O2)

  • Ahmed, Syed Rahin;Koh, Kwang-Nak;Kang, Nam-Lyong;Lee, Jae-Beom
    • Bulletin of the Korean Chemical Society
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    • v.33 no.5
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    • pp.1608-1612
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    • 2012
  • Singlet oxygen ($^1O_2$) is an important species for oxidation in biological processes. $^1O_2$ is implicated in the genotoxic effect, and plays an important role in the cell-signaling cascade and in the induction of gene expression. However, the rapid detection of $^1O_2$ in biological environments with sufficient specificity and sensitivity is hampered by its extremely low emission probability. Here, a layer-by-layer (LbL) film of CdTe quantum dots (QDs), polymers, and ascorbate have been designed as a rapid, highly selective, and sensitive fluorescence probe for $^1O_2$ detection. Upon reaction with $^1O_2$, the probe exhibits a strong photoluminescence (PL) response even at trace levels. This remarkable PL change should enable the probe to be used for $^1O_2$ detection in many chemical and biological systems and as an environmental sensor.

Development of Differential Media and Multiplex PCR Assays for the Rapid Detection of Listeria monocytogenes (Listeria monocytogenes의 신속검출을 위한 선택배지 및 multiplex PCR 기법 개발)

  • Jung, Byeong-yeal;Lim, Hyun-sook;Jung, Suk-chan
    • Korean Journal of Veterinary Research
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    • v.43 no.2
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    • pp.231-237
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    • 2003
  • Listeria (L.) monocytogenes in samples could not be detected occasioally by faster growth of other Listeria spp. especially L. innocua. The aim of this study was to develop the differential media and multiplex polymerase chain reaction (PCR) assays for the rapid detection of L. monocytogenes. L. monocytogenes colonies were characterized by their ${\beta}$-hemolysis with fluorescence under 366 nm UV light on the Listeria hemolysis agar (LHA). L. innocua, a species commonly present in foods, did not produce ${\beta}$-hemolysis on LHA. Therefore, one or more colonies of L. monocytogenes were easily distinguished from large populations of L. innocua. The multiplex PCR assays were developed to distinguish from L. monocytogenes and other Listeria spp. with two pairs of primers. The primers were designed in 16S rRNA and listeriolysin O gene for specific amplification of all members of the genus Listeria and L. monocytogenes, respectively. The multiplex PCR assays produced 560 and 938 bp products in L. monocytogenes; only 938 bp products in the genus Listeria. The multiplex PCR assays could detect as little as 50 pg of L monocytogenes DNA. These results indicated that the differential media and multiplex PCR assays might be useful diagnostic tools for the rapid detection of L. monocytogenes.

Detection of Red Tide Patches using AVHRR and Landsat TM data (AVHRR과 Landsat TM 자료를 이용한 적조 패취 관측)

  • Jeong, Jong-Chul
    • Journal of Environmental Impact Assessment
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    • v.10 no.1
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    • pp.1-8
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    • 2001
  • Detection of red tides by satellite remote sensing can be done either by detecting enhanced level of chlorophyll pigment or by detecting changes in the spectral composition of pixels. Using chlorophyll concentration, however, is not effective currently due to the facts: 1) Chlorophyll-a is a universal pigment of phytoplankton, and 2) no accurate algorithm for chlorophyll in case 2 water is available yet. Here, red band algorithm, classification and PCA (Principal Component Analysis) techniques were applied for detecting patches of Cochlodinium polykrikoides red tides which occurred in Korean waters in 1995. This dinoflagellate species appears dark red due to the characteristic pigments absorbing lights in the blue and green wavelength most effectively. In the satellite image, the brightness of red tide pixels in all the three visible bands were low making the detection difficult. Red band algorithm is not good for detecting the red tide because of reflectance of suspended sediments. For supervised classification, selecting training area was difficult, while unsupervised classification was not effective in delineating the patches from surrounding pixels. On the other hand, PCA gave a good qualitative discrimination on the distribution compared with actual observation.

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Fluorescence Characteristic Spectra of Domestic Fuel Products through Laser Induced Fluorescence Detection

  • Wu, Ting-Nien;Chang, Shui-Ping;Tsai, Wen-Hsien;Lin, Cian-Yi
    • Journal of Soil and Groundwater Environment
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    • v.19 no.5
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    • pp.18-25
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    • 2014
  • Traditional investigation procedures of soil and groundwater contamination are followed by soil gas sampling, soil sampling, groundwater sampling, establishment of monitoring wells, and groundwater monitoring. It often takes several weeks to obtain the analysis reports, and sometimes, it needs supplemental sampling and analysis to delineate the polluted area. Laser induced fluorescence (LIF) system is designed for the detection of free-phase petroleum pollutants, and it is suitable for on-site real-time site investigation when coupling with a direct push testing tool. Petroleum products always contain polycyclic aromatic hydrocarbon (PAH) compounds possessing fluorescence characteristics that make them detectable through LIF detection. In this study, LIF spectroscopy of 5 major fuel products was conducted to establish the databank of LIF fluorescence characteristic spectra, including gasoline, diesel, jet fuel, marine fuel and low-sulfur fuel. Multivariate statistical tools were also applied to distinguish LIF fluorescence characteristic spectra among the mixtures of selected fuel products. This study successfully demonstrated the feasibility of identifying fuel species based on LIF characteristic fluorescence spectra, also LIF seemed to be uncovered its powerful ability of tracing underground petroleum leakages.

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.

Analysis of Heterocyclic Amines in Human Urine Using Multiple Solid-Phase Extraction by Liquid Chromatography/Mass Spectrometry

  • Cha, Hyun-Jeong;Kim, Nam-Hee;Jeong, Eun-Kyung;Na, Yun-Cheol
    • Bulletin of the Korean Chemical Society
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    • v.31 no.8
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    • pp.2322-2328
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    • 2010
  • A multiple solid-phase extraction (SPE) method was used with liquid chromatography, coupled with mass spectrometry (LC/MS), for the analysis of heterocyclic amines (HCAs) in human urine. Separation efficiencies based on the pH of the mobile phase and the types of columns were compared. An amide column showed better baseline separation and narrower HCA peak widths at pH 5.0 for the mobile phase than a $C_8$ column. Each SPE step, HLB, MCX, and HybridSPE, was optimized by controlling the pH conditions. The combined method with the three SPEs effectively removed interfering species that cause ion-suppression during HCA detection. Validation of the method, performed with SIM and SRM detection, showed correlation coefficients above 0.991 in the range 0.3 - 16.7 ng/mL. Recovery rates were 45.4 - 97.3% on the $C_8$ column and 71.8 - 101.4% on the amide column, and method detection limits were 0.11 - 0.65 ng/mL on the $C_8$ column and 0.12 - 0.48 ng/mL on the amide column. This method using multiple SPEs offers significant benefits for high-throughput determination of HCAs in urine.

Loop-Mediated Isothermal Amplification for the Detection of Xanthomonas arboricola pv. pruni in Peaches

  • Li, Weilan;Lee, Seung-Yeol;Back, Chang-Gi;Ten, Leonid N.;Jung, Hee-Young
    • The Plant Pathology Journal
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    • v.35 no.6
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    • pp.635-643
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    • 2019
  • To detect Xanthomonas arboricola pv. pruni, a loopmediated isothermal amplification (LAMP) detection method were developed. The LAMP assay was designed to test crude plant tissue without pre-extraction, or heating incubation, and without advanced analysis equipment. The LAMP primers were designed by targeting an ABC transporter ATP-binding protein, this primer set was tested using the genomic DNA of Xanthomonas and non-Xanthomonas strains, and a ladder product was generated from the genomic DNA of X. arboricola pv. pruni strain but not from 12 other Xanthomonas species strains and 6 strains of other genera. The LAMP conditions were checked with the healthy leaves of 31 peach varieties, and no reaction was detected using either the peach leaves or the peach DNA as a template. Furthermore, the high diagnostic accuracy of the LAMP method was confirmed with 13 X. arboricola pv. pruni strains isolated from various regions in Korea, with all samples exhibiting a positive reaction in LAMP assays. In particular, the LAMP method successfully detected the pathogen in diseased peach leaves and fruit in the field, and the LAMP conditions were proven to be a reliable diagnostic method for the specific detection and identification of X. arboricola pv. pruni in peach orchards.

A Simultaneous Analytical Method for Duplex Identification of Porcine and Horse in the Meat Products by EvaGreen based Real-time PCR

  • Sakalar, Ergun;Ergun, Seyma Ozcirak;Akar, Emine
    • Food Science of Animal Resources
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    • v.35 no.3
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    • pp.382-388
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    • 2015
  • A duplex real-time polymerase chain reaction (PCR) based assay for the detection of porcine and horse meat in sausages was designed by using EvaGreen fluorescent dye. Primers were selected from mitochondrial 12S rRNA and 16S rRNA genes which are powerful regions for identification of horse and porcine meat. DNA from reference samples and industrial products was successfully extracted using the GIDAGEN® Multi-Fast DNA Isolation Kit. Genomes were identified based on their specific melting peaks (Mp) which are 82.5℃ and 78℃ for horse and porcine, respectively. The assay used in this study allowed the detection of as little as 0.0001% level of horse meat and 0.001% level of porcine meat in the experimental admixtures. These findings indicate that EvaGreen based duplex realtime PCR is a potentially sensitive, reliable, rapid and accurate assay for the detection of meat species adulterated with porcine and horse meats.

Multiplex RT-PCR Assay for the Detection of Apple stem grooving virus and Apple chlorotic leaf spot virus in Infected Korean Apple Cultivars

  • Park, Hong-Lyeol;Yoon, Jae-Seung;Kim, Hyun-Ran;Baek, Kwang-Hee
    • The Plant Pathology Journal
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    • v.22 no.2
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    • pp.168-173
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    • 2006
  • To develop the diagnostic method for the viral infection in apple, the partial genes corresponding to the N-terminal region of RNA polymerase of Apple stem grooving virus (ASGV) and coat protein of Apple chlorotic leaf spot virus (ACLSV) were characterized from the infected apple cultivars in Korea. Based on the nucleotide sequences of the characterized partial genes, the virus gene-specific primers were designed for the detection of ASGV and ACLSV infected in species of Malus. The RT-PCR using the primers for the genes of ASGV and ACLSV successfully gave rise to 404 and 566 bp DNA fragments, respectively. Using those viral gene-specific primers, the multiplex RT-PCR assays were also established to diagnose the mixed infection by ASGV and ACLSV simultaneously. Furthermore, the control primers, which have to be included for the RT-PCR as an internal control, were designed using the nucleotide sequence of the gene encoding elongation factor $1{\alpha}(EF1{\alpha})$. This multiplex RT-PCR including the control primers provides more reliable, rapid and sensitive assay for the detection of ASGV and ACLSV infected in Korean apple cultivars.