• 식물병연구
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• 제27권1호
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• pp.38-44
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• 2021

Xylella fastidiosa is the most damaging pathogen in many parts of the world. To increase diagnostic capability of X. fastidiosa in the field, the loop-mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR) assay were developed to mqsA gene of citrate-synthase (XF 1535) X. fastidiosa and evaluated for specificity and sensitivity. Both assays were more robust than current published tests for detection of X. fastidiosa when screened against 16 isolates representing the four major subgroups of the bacterium from a range of host species. No cross reaction with DNA from healthy hosts or other species of bacteria has been observed. The LAMP and PCR assays could detect 10-4 pmol and 100 copies of the gene, respectively. Hydroxynaphthol blue was evaluated as an endpoint detection method for LAMP. There was a significant color shift that signaled the existence of the bacterium when at least 100 copies of the target template were present.

• 센서학회지
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• 제31권4호
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• pp.187-193
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• 2022

The accurate detection of environmental and biomarker gas species has attracted increasing attention due to their broad applications, such as air quality monitoring, disease diagnosis, and explosive chemicals detection. To accurately detect target gas species using chemiresistive gas sensors, using nanocatalysts on semiconducting metal oxides (SMOs) is considered the most promising approach. This review summarizes recent studies on methods for nanocatalysts functionalization on SMOs to achieve the highly selective gas sensors. To this end, we discuss various nanocatalyst decorated metal oxide-based chemiresistive gas sensors and provide an insight to construct highly accurate gas sensors.

• Parasites, Hosts and Diseases
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• 제21권2호
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• pp.187-192
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• 1983

In order to clarify infestation Pattern for the encysted larvae of digenetic trematodes from fresh-water fishes, this survey was carried out from March to September, 1983. A total of 380 fishes of 32 species wore collected with netting at the three reaches, upper, middle and lower in Mangyeong riverside area. After the fishes were dissected into small scraps, they were pressed under cover glass and examined-for the presence of those of digenetic trematodes with a microscope. The resillts obtailled were as follows; Out of a total of 380 knishes inspected, 320 fishes(84%) from 31 species were found positive with, digenetic trematode metacercariae; more than 10 species of the metacercariae Ivere detected in Pseuderasbora parva; .Gnnthepegen mtajimae, Microphysogokio yaluensis, Cultriculus eigenmanni and Gnnthopogon coreanus (more than 8 species) ; Aphyocypris chinensis (8 species) and etc. respectively. Clonorchis sinensis metacercariae were found positive from 93 fishes (25%) from 12 species and detection rates in other species of digenetic trematode metacercariae from various fishes were; Exorchis oviformis, 261 fisles (57%) from 28 species; Cyathocotyle criensalis, 47 fishcs (12%) from 12 species; Metorchis orientalis, 21 fishes (6%) from 12 species; Metagonimus yokogawai, 164 fishes (43%) from 26 species; Pseudesorchis major, 71 fishes (19%) from 18 species; Metacercaria haiegawai, 77 fishes (20%) from 25 species; Centrocestus armatus, 24 fishes (6%) from 7 species; Echinochasmus japonicus, 2 fishes (0.5%) from 2 species, and unidentified species, 34 fishes (9%) from 15 species respectively. The sums of average number of the encysted larvae of all species found in fish body/gram showed 83 in P.parva, Cobitis taenia (74.2), A. chinensis (28.5), Pseudoperilampus uyekii (26.6), G. majimae (19.6) and etc. respectively and the average peak number of each metacercaria in fish body/gram showed 21.7 C. sinensis, 245. ovifcrmis, 15, 3 M, crientalis and 6.1 5. japonicus in P. parva. 42.7 C. orientalis and 25.1 M. yohogawai in C. taenia; 8.3 C. armatus and 8.3 M. hasegawai in P. uyekii. 6.3 P. major in Carassius carassius, and 2.9 unidentified species in G, niajimae respectively.

• Food Science and Biotechnology
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• 제17권2호
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• pp.357-361
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• 2008

One of the major allergenic proteins in common buckwheat (Fagopyrum elculentum) was found to be a BW10KD. In this work, allergenic BW10KD genomic DNAs from the native common buckwheat 'Pyeongchang' and Tartarian buckwheat 'Clfa47' were cloned by polymerase chain reaction (PCR), and their nucleotide sequences were determined. In addition, a novel PCR assay targeting the allergenic BW10KD gene was developed to detect and differentiate both buckwheat species in food. The nucleotide sequences of the BW10KD genomic DNA from 'Pyeongchang' and 'Clfa47' were 94% identical. Base differences in the nucleotide sequences of the BW10KD genes are probably useful as a molecular marker for species-specific identification. The 'Pyeongchang'-specific primer set 154PF/400PR and the 'Clfa47'-specific primer set 154DF/253DR generated 247 and 100 bp fragments in singleplex PCR, respectively. A duplex PCR assay with 2 species-specific primer sets simultaneously differentiated the 'Pyeongchang' and 'Clfa47' in a single reaction. The PCR assay also successfully allowed for the rapid detection of buckwheat ingredients in foods.

• Asian-Australasian Journal of Animal Sciences
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• 제16권3호
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• pp.425-429
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• 2003

Staphylococcus aureus is a major pathogen for cattle, causing various forms of subclinical and clinical mastitis and could be a causative agent of food poisoning, it produces various superantigenic exotoxins which have a great public health significance. A total of 72 S. aureus clinical isolates from dairy farms located in Kyunggi Province Korea were examined for the species identification by biochemical method, and for the detection of $\beta$-lactamase, enterotoxin and other exotoxins genes by PCR. The results of species identification by biochemical method agreed with those of PCR done with species specific primer STA-AU. $\beta$-lactamase is an enzyme closely associated with the resistance to antibiotic penicillin, which is an important means of treatment of mastitis, all the isolates were positive for the presence of genes encoding $\beta$-lactamase, which were reproduced in penicillin susceptibility disc assay. Six types of toxin genes, Staphylococcal enterotoxin (SE)A, SEB, SEC, SEE, toxic shock syndrome toxin (TSST-1) and exfoliative toxin A (ET A) were detected in 72 isolates by PCR associated genotypic method in this study, none of the isolates carried the genes for enterotoxin D (SED) and exfoliative toxin B (ETB). The occurrence rate of exotoxin genes rated as 12.5%, and the precision of the PCR identification results has been confirmed using the reference strains.

• The Plant Pathology Journal
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• 제27권2호
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• pp.120-127
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• 2011

While many isolates of Alternaria alternata are common saprophytes on trees and shrubs, this study clearly demonstrated that A. alternata is a primary pathogen in lilac (Syringa sp.), causing a leaf-blight that affects different Syringa species. Isolates of Alternaria sp. were collected from leaf blight samples of lilacs in the field. The internal transcribed spacer (ITS) region and morphological characterization were used to identify lilac blight pathogen. Based on 100% ITS nucleotide sequence identities to the Alternaria genus in the GenBank and morphological features, these isolates were identified as A. alternata. Disease symptoms were reproduced in lilac plants inoculated with A. alternata mycelial plugs and sprayed with a fungus-free culture filtrate, indicating that pathogenesis in lilac involves secondary metabolites or toxins. Diagnostic primers were developed to detect Alternaria sp. and A. alternata in lilac leaf blight based on ITS region and four known genes associated with pathogenesis in A. alternata: mixed-linked glucanase precursor, endopolygalacturonase, hsp70, and histone genes. The results from our study indicated A. alternata is a primary pathogen in lilac leaf blight, and these diagnostic primers can be used as a tool for the fast detection of A. alternata associated with lilac leaf blight.

• 한국동물위생학회지
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• 제37권4호
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• pp.247-252
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• 2014

The economic impact of swine mycoplasma infection is high. An accurate diagnosis is often difficult and time consuming. We report the development and validation of an effective multiplex polymerase chain reaction (PCR) assay that detects Mycoplasma (M.) hyopneumoniae and M. hyorhinis. The multi detection of M. hyopneumoniae and M. hyorhinis primer set were employed to detect mycoplasma species and typing of the species was performed on the basis of sequence analysis of the PCR product. The target nucleic acid fragments were specifically amplified by M. hyopneumoniae and M. hyorhinis PCR with 16S ribosomal DNA primers. Single and mixed Mycoplasma species DNA templates were used to evaluate the specificity of the multiplex assay. The corresponding specific DNA products were amplified for each pathogen. The multiplex PCR assay provides a novel tool for simultaneous detection and differentiation of M. hyopneumoniae and M. hyorhinis.

• Bulletin of the Korean Chemical Society
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• 제25권1호
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• pp.73-78
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• 2004

A highly sensitive detecting method has been developed for determining part per billion of sulfur in $H_2S$/Ar plasma. The method is based on the excitation of Ar/$H_2S\;or\;Ar/H_2S/O_2$ mixture in hollow cathode glow discharge sustained by radiofrequency (RF) or 60 Hz AC power and the spectroscopic measurement of the intensity of emission lines from electronically excited $S_2^*\;or\;SO_2^*$ species, respectively. The RF or AC power needed for the excitation did not exceed 30 W at a gas pressure maintained at several mbar. The emission intensity from the $SO_2^*$ species showed excellent linear response to the sulfur concentration ranging from 5 ppbv, which correspond to S/N = 5, to 500 ppbv. But the intensity from the $S_2^*$ species showed a linear response to the $H_2S$ only at low flow rate under 20 sccm (mL/min) of the sample gas. Separate experiments using $SO_2$ gas as the source of sulfur demonstrated that the presence of $O_2$ in the argon plasma is essential for obtaining prominent $SO_2^*$ emission lines.

• Journal of Microbiology and Biotechnology
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• 제17권3호
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• pp.481-489
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• 2007

[ $\alpha$ ]-L-Arabinofuranosidases (AFases; EC 3.2.1.55) are exo-type enzymes, which hydrolyze terminal nonreducing arabinose residues from various polysaccharides such as arabinan and arabinoxylan. Genome-wide BLAST search showed that various bacterial strains possess the putative AFase genes with well-conserved motif sequences at the nucleotide and amino acid sequence levels. In this study, two sets of degenerate PCR primers were designed and tested to detect putative AFase genes, based on their three highly conserved amino acid blocks (PGGNFV, GNEMDG; and DEWNVW). Among 20 Bacillus-associated species, 13 species were revealed to have putative AFase genes in their genome and they share over 67% of amino acid identities with each other. Based on the partial sequence obtained from an isolate, an AFase from Geobacillus sp. was cloned and expressed in E. coli. Enzymatic characterization has verified that the resulting enzyme corresponds to a typical AFase. Accordingly, degenerate PCR primers developed in this work can be used for fast, easy, and specific detection and isolation of putative AFase genes from bacterial cells.

• Journal of Microbiology and Biotechnology
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• 제12권6호
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• pp.989-992
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• 2002

Sweet persimmon (Diospyros kaki Thunb.) is widely cultivated in the southern part of Korea and its cultivation is increasing. However, anthracnose disease caused by Colletotricuhum species is one of the major hinderances to the cultivation and production of sweet persimmon. Therefore, in the current study, PCR was used to specifically detect Colletotrichum spp., based on the sequences of the ITS II regions in the rDNA. Using the sequence data, CO-1 was designated to detect Colletotrichum together the with ITS 4 primer. The result showed that a single segment of ca. 500 bp was observed only in Colletotrichum, but not in any other fungal and bacterial isolates. The annealing temperatures and template DNA quantites were also investigated to identify optimal conditions for detection. Using these species-specific primers, a unique band was obtained at annealing temperatures ranging from $55^{\circ}C\;and\;61^{\circ}C$ and template DNA levels from 10 pg- $10{\mu}g$.