• Title/Summary/Keyword: Species detection

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Detection of Aspergillus, Penicillium, and Fusarium Species by Sandwich Enzyme-Linked Immunosorbent Assay Using Mixed Monoclonal Antibodies

  • Kwak, Bo-Yeon;Kwon, Byung-Joon;Kweon, Chang-Hee;Shon, Dong-Hwa
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.385-389
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    • 2004
  • The antibody-mix sandwich enzyme-linked immunosorbent assay (Ab-mix sELlS A) system was developed in order to simultaneously detect the extracellular polysaccharide (FPS) of Aspergillus, Penicillium, or Fusarium species using one detection system. The detection limit and detection range of the Ab-mix sELISA towards EPS of Penicilliun citrinum were not changed, and those towards Fusarium moniliforme EPS were changed a little compared to that of individual sandwich ELISA [9, 10]. The fungal culture filtrates of Aspergillus and Penicillium species showed nearly similar reactivity towards Ab-mix sELISA as that of sELISA using the MAb lB8 alone [9]. Also, the fungal culture filtrates of Fusarium species showed nearly the same reactivity towards Ab-mix sELISA as that of sELISA using the MAb lB8 alone [10]. Thus, this ELISA system showed that the three genera of molds, Aspergillus, Penicillium, or Fusarium, which are three major important molds producing mycotoxins in food or agricultural commodities, could be detected at the same time, using one detection system.

Microcystin Detection Characteristics of Fluorescence Immunochromatography and High Performance Liquid Chromatography

  • Pyo, Dong-Jin;Park, Geun-Young;Choi, Jong-Chon;Oh, Chang-Suk
    • Bulletin of the Korean Chemical Society
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    • v.26 no.2
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    • pp.268-272
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    • 2005
  • Different detection characteristics of fluorescence immunochromatography method and high performance liquid chromatography (HPLC) method for the analysis of cyanobacterial toxins were studied. In particular, low and high limits of detection, detection time and reproducibility and detectable microcystin species were compared when fluorescence immunochromatography method and high performance liquid chromatography method were applied for the detection of microcystin (MC), a cyclic peptide toxin of the freshwater cyanobacterium Microcystis aeruginosa. A Fluorescence immunochromatography assay system has the unique advantages of short detection time and low detection limit, and high performance liquid chromatography detection method has the strong advantage of individual quantifications of several species of microcystins.

Development of a Species-specific PCR Assay for Three Xanthomonas Species, Causing Bulb and Flower Diseases, Based on Their Genome Sequences

  • Back, Chang-Gi;Lee, Seung-Yeol;Lee, Boo-Ja;Yea, Mi-Chi;Kim, Sang-Mok;Kang, In-Kyu;Cha, Jae-Soon;Jung, Hee-Young
    • The Plant Pathology Journal
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    • v.31 no.3
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    • pp.212-218
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    • 2015
  • In this study, we developed a species-specific PCR assay for rapid and accurate detection of three Xanthomonas species, X. axonopodis pv. poinsettiicola (XAP), X. hyacinthi (XH) and X. campestris pv. zantedeschiae (XCZ), based on their draft genome sequences. XAP, XH and XCZ genomes consist of single chromosomes that contain 5,221, 4,395 and 7,986 protein coding genes, respectively. Species-specific primers were designed from variable regions of the draft genome sequence data and assessed by a PCR-based detection method. These primers were also tested for specificity against 17 allied Xanthomonas species as well as against the host DNA and the microbial community of the host surface. Three primer sets were found to be very specific and no amplification product was obtained with the host DNA and the microbial community of the host surface. In addition, a detection limit of $1pg/{\mu}l$ per PCR reaction was detected when these primer sets were used to amplify corresponding bacterial DNAs. Therefore, these primer sets and the developed species-specific PCR assay represent a valuable, sensitive, and rapid diagnostic tool that can be used to detect three specific pathogens at early stages of infection and may help control diseases.

Phylogenetic Analysis and Rapid Detection of Genus Phellinus using the Nucleotide Sequences of 18S Ribosomal RNA

  • Nam, Byung-Hyouk;Lee, Jae-Yun;Kim, Gi-Young;Jung, Heon-Ho;Park, Hyung-Sik;Kim, Cheng-Yun;Jo, Wol-Soon;Jeong, Soo-Jin;Lee, Tae-Ho;Lee, Jae-Dong
    • Mycobiology
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    • v.31 no.3
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    • pp.133-138
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    • 2003
  • Analysis of phylogenetic relationship was performed among Phellinus species based on 18S ribosomal subunit sequence data. Twenty-five strains of 19 Phellinus species including P. linteus were examined in this study. Regions of 18S ribosomal subunit were very conserved, but some variable regions between Phellinus species were observed. The species-specific detection primers, modified by 2 or 3 nucleotides in sense primer were designed based on 18S ribosomal DNA(rDNA) sequence data. The 210 by PCR bands were detected with annealing temperature $48^{\circ}C$. The 18S 2F-18S 4R detection primer set distinguished P. linteus from various Phellinus species but some species like P. baumii, P. weirianius, P. rhabarberinus and P. pomaceus also had weak reactivity on this primer set. The 18S 3F-18S 4R primer set distinguished only P. linteus from various Phellinus species, although sensitivity with this primer set was lower than that of 18S 2F-18 4R primer set. These primer sets would be useful for the detection of only P. linteus among unknown Phellinus species rapidly.

A fast and reliable polymerase chain reaction method based on short interspersed nuclear elements detection for the discrimination of buffalo, cattle, goat, and sheep species in dairy products

  • Cosenza, Gianfranco;Iannaccone, Marco;Gallo, Daniela;Pauciullo, Alfredo
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.6
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    • pp.891-895
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    • 2019
  • Objective: Aim of present study was the set up of a fast and reliable protocol using species-specific markers for the quali-quantitative analysis of DNA and the detection of ruminant biological components in dairy products. For this purpose, the promoter of the gene coding for the ${\alpha}$-lactoalbumin (LALBA) was chosen as possible candidate for the presence of short interspersed nuclear elements (SINEs). Methods: DNA was isolated from somatic cells of 120 individual milk samples of cattle (30), Mediterranean river buffalo (30), goat (30), and sheep (30) and the gene promoter region (about 600/700 bp) of LALBA (from about 600 bp upstream of exon 1) has been sequenced. For the development of a single polymerase chain reaction (PCR) protocol that allows the simultaneous identification of DNA from the four species of ruminants, the following internal primers pair were used: 5'-CACTGATCTTAAAGCTCAGGTT-3' (forward) and 5'-TCAGA GTAGGCCACAGAAG-3' (reverse). Results: Sequencing results of LALBA gene promoter region confirmed the presence of SINEs as monomorphic "within" and variable in size "among" the selected species. Amplicon lengths were 582 bp in cattle, 592 bp in buffalo, 655 in goat and 729 bp in sheep. PCR specificity was demonstrated by the detection of trace amounts of species-specific DNA from mixed sources ($0.25ng/{\mu}L$). Conclusion: We developed a rapid PCR protocol for the quali-quantitative analysis of DNA and the traceability of dairy products using a species-specific marker with only one pair of primers. Our results validate the proposed technique as a suitable tool for a simple and inexpensive (economic) detection of animal origin components in foodstuffs.

Direct Determination of Total Arsenic and Arsenic Species by Ion Chromatography Coupled with Inductively Coupled Plasma Mass Spectrometry

  • Nam, Sang-Ho;Kim, Jae-Jin;Han, Soung-Sim
    • Bulletin of the Korean Chemical Society
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    • v.24 no.12
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    • pp.1805-1808
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    • 2003
  • The simultaneous determination of As(III), As(V), and DMA has been performed by ion chromatography (IC) coupled with inductively coupled plasma-mass spectrometry (ICP-MS). The separation of the three arsenic species was achieved by an anionic separator column (AS 7) with an isocratic elution system. The separated species were directly detected by ICP-MS as an element-selective detection method. The IC-ICP-MS technique was applied for the determination of arsenic species in a NIST SRM 1643d water sample. An As(III) only was detected in the sample. The detection limits of As(III), As(V) and DMA were 0.31, 0.45, and 2.09 ng/mL, respectively. It was also applied for the determination of arsenic species in a human urine obtained by a volunteer, and three arsenic species were identified. The determination of total As in human urines that were obtained from 25 volunteers at the different age was also carried out by ICP-MS.

Multiplex PCR Assay for Simultaneous Detection of Korean Quarantine Phytoplasmas

  • Kim, Young-Hwan;Win, Nang Kyu;Back, Chang-Gi;Yea, Mi-Chi;Yim, Kyu-Ock;Jung, Hee-Young
    • The Plant Pathology Journal
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    • v.27 no.4
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    • pp.367-371
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    • 2011
  • Multiplex PCR assays were developed for the simultaneous detection of ten important Korean quarantine phytoplasmas. The species-specific primers were designed based on ribosomal protein, putative preprotein translocase Y, immunodominant protein, elongation factor TU, chaperonin protein and the 16S rRNA genes of 'Candidatus (Ca.) Phytoplasma' species. Three main primer sets were prepared from ten designed primer pairs to limit nonspecific amplification as much as possible. The multiplex PCR assay using the three primer sets successfully amplified the correct conserved genes for each 'Ca. Phytoplasma' species. In addition, ten important 'Ca. Phytoplasma' species could be easily determined by recognizing band patterns specific for each phytoplasma species from three primer sets. Moreover, a high sensitivity of multiplex PCR for each primer set was observed for samples containing a low DNA concentration (10 ng/${\mu}l$). This study provides the useful multiplex PCR assay as a convenient method to detect the presence of ten important quarantine phytoplasmas in Korea.

Development of Urban Wildlife Detection and Analysis Methodology Based on Camera Trapping Technique and YOLO-X Algorithm (카메라 트래핑 기법과 YOLO-X 알고리즘 기반의 도시 야생동물 탐지 및 분석방법론 개발)

  • Kim, Kyeong-Tae;Lee, Hyun-Jung;Jeon, Seung-Wook;Song, Won-Kyong;Kim, Whee-Moon
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.26 no.4
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    • pp.17-34
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    • 2023
  • Camera trapping has been used as a non-invasive survey method that minimizes anthropogenic disturbance to ecosystems. Nevertheless, it is labor-intensive and time-consuming, requiring researchers to quantify species and populations. In this study, we aimed to improve the preprocessing of camera trapping data by utilizing an object detection algorithm. Wildlife monitoring using unmanned sensor cameras was conducted in a forested urban forest and a green space on a university campus in Cheonan City, Chungcheongnam-do, Korea. The collected camera trapping data were classified by a researcher to identify the occurrence of species. The data was then used to test the performance of the YOLO-X object detection algorithm for wildlife detection. The camera trapping resulted in 10,500 images of the urban forest and 51,974 images of green spaces on campus. Out of the total 62,474 images, 52,993 images (84.82%) were found to be false positives, while 9,481 images (15.18%) were found to contain wildlife. As a result of wildlife monitoring, 19 species of birds, 5 species of mammals, and 1 species of reptile were observed within the study area. In addition, there were statistically significant differences in the frequency of occurrence of the following species according to the type of urban greenery: Parus varius(t = -3.035, p < 0.01), Parus major(t = 2.112, p < 0.05), Passer montanus(t = 2.112, p < 0.05), Paradoxornis webbianus(t = 2.112, p < 0.05), Turdus hortulorum(t = -4.026, p < 0.001), and Sitta europaea(t = -2.189, p < 0.05). The detection performance of the YOLO-X model for wildlife occurrence was analyzed, and it successfully classified 94.2% of the camera trapping data. In particular, the number of true positive predictions was 7,809 images and the number of false negative predictions was 51,044 images. In this study, the object detection algorithm YOLO-X model was used to detect the presence of wildlife in the camera trapping data. In this study, the YOLO-X model was used with a filter activated to detect 10 specific animal taxa out of the 80 classes trained on the COCO dataset, without any additional training. In future studies, it is necessary to create and apply training data for key occurrence species to make the model suitable for wildlife monitoring.

Identification and Detection of Streptococcus anginosus Using Species-Specific 16S rDNA Primers

  • Cho, Ji-Sun;Yoo, So-Young;Kim, Hwa-Sook;Hwang, Ho-Keel;Min, Jeong-Beom;Kim, Byung-Hoon;Baek, Dong-Heon;Shin, Hwan-Seon;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.31 no.1
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    • pp.11-14
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    • 2006
  • This study was undertaken to develop PCR primers for the identification and detection of Streptococcus anginosus using species-specific forward and reverse primers. These primers targeted the variable regions of the 16S ribosomal RNA coding gene(rDNA). The primer specificity was tested against 12 S. anginosus strains and 6 different species(10 strains) of oral bacteria. The primer sensitivity was determined by testing serial dilutions of the purified genomic DNA of S. anginosus ATCC $33397^T$. The data showed that species-specific amplicons were obtained from all the S. anginosus strains tested, but not in the six other species. The PCR could detect as little as 0.4pg of the chromosomal DNA from S. anginosus. This suggests that the PCR primers are highly sensitive and applicable to the detection and identification of S. anginosus.

Development of a Rapid and Accurate Identification Method for Citrobacter Species Isolated from Pork Products Using a Matrix-Assisted Laser-Desorption Ionization Time-of-Flight Mass Spectrometry (MALDITOF MS)

  • Kwak, Hye-Lim;Han, Sun-Kyung;Park, Sunghoon;Park, Si Hong;Shim, Jae-Yong;Oh, Mihwa;Ricke, Steven C.;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.25 no.9
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    • pp.1537-1541
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    • 2015
  • Previous detection methods for Citrobacter are considered time consuming and laborious. In this study, we have developed a rapid and accurate detection method for Citrobacter species in pork products, using matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). A total of 35 Citrobacter strains were isolated from 30 pork products and identified by both MALDI-TOF MS and 16S rRNA gene sequencing approaches. All isolates were identified to the species level by the MALDI-TOF MS, while 16S rRNA gene sequencing results could not discriminate them clearly. These results confirmed that MALDITOF MS is a more accurate and rapid detection method for the identification of Citrobacter species.