• Molecules and Cells
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• v.24 no.3
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• pp.329-337
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• 2007

An enrichment semi-nested PCR procedure was developed for detection of Clostridium botulinum types A, B, E, and F. It was applied to sediment samples to examine the prevalence of C. botulinum in the Korean environment. The first pair of primers for the semi-nested PCR was designed using a region shared by the types A, B, E, and F neurotoxin gene sequences, and the second round employed four nested primers complementary to the BoNT/A, /B, /E, and /F encoding genes for simultaneous detection of the four serotypes. Positive results were obtained from the PCR analysis of five of 44 sediments (11%) collected from Yeong-am Lake in Korea; all were identified as deriving from type B neurotoxin (bontb) genes. Two of the C. botulinum type B organisms were isolated, and their bontb genes sequenced. The deduced amino acid sequences of BoNT/B showed 99.5 and 99.8% identity with the amino acid sequence of accession no. AB084152. Our data suggest that semi-nested PCR is a useful tool for detecting C. botulinum in sediments, and renders it practicable to conduct environmental surveys.

• Journal of Microbiology and Biotechnology
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• v.23 no.7
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• pp.1041-1045
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• 2013

The xTAG$^{(R)}$ Gastrointestinal Pathogen Panel (GPP) is a multiplexed molecular test for 15 gastrointestinal pathogens. The sensitivity and specificity of this test were assessed in 901 stool specimens collected from pediatric and adult patients at four clinical sites. A combination of conventional and molecular methods was used as comparator. Sensitivity could be determined for 12 of 15 pathogens and was 94.3% overall. The specificity across all 15 targets was 98.5%. Testing for the pathogen identified was not requested by the physician in 65% of specimens. The simultaneous detection of these 15 pathogens can provide physicians with a more comprehensive assessment of the etiology of diarrheal disease.

• Bulletin of the Korean Chemical Society
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• v.27 no.4
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• pp.479-483
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• 2006

We developed a microfluidic immunoassay platform for the detection of various analytes such as bacterial pathogen by packing antibody-immobilized glass beads in spatially-isolated microchambers on a microfluidic device. Primary amines of antibody were covalently conjugated to carboxyl-terminated glass beads previously treated with aminosilane followed by glutaraldehyde. Through this covalent binding, up to 905 $\mu$g immunoglobulin G (IgG) per gram of glass beads was immobilized. For application, glass beads attaching antibody specific to Escherichia coli O157:H7, a foodborne pathogen, were packed into a microfluidic device and used for the detection of the serotype. This prototype immunoassay device can be used for the simultaneous detection of multiple analytes by sequentially packing different-sized glass beads attaching different antibody in discrete microchambers on a single microfluidic device.

• The Plant Pathology Journal
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• v.27 no.2
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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.

• Journal of Microbiology and Biotechnology
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• v.16 no.5
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• pp.740-747
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• 2006

Amplification by universal consensus sequences in pathogenic bacterial DNA would allow rapid identification of pathogenic bacteria, and amplification of genus-specific and species-specific sequences of pathogenic bacterial DNA might be used for genotyping at the genus and species levels. For design of probes for molecular diagnostics, several tools are available as stand-alone programs or as Web application. However, since most programs can design only a few probe sets at one time, they are not suitable for large-scale and automatic probes design. Therefore, for high-throughput design of specific probes in diagnostic array development, an automated design tool is necessary. Thus, we developed a Web-based automatic system for design of genus-specific and species-specific probes for pathogen detection. The system is available at http://www.miprobe.com.

• The Plant Pathology Journal
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• v.32 no.1
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• pp.53-57
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• 2016

Major diseases in grafted cacti have been reported and Fusarium oxysporum, Bipolaris cactivora, Phytophthora spp. and Collectotrichum spp. are known as causal pathogens. These pathogens can lead to plant death after infection. Therefore, some European countries have quarantined imported cacti that are infected with specific fungal pathogens. Consequently, we developed PCR detection methods to identify four quarantined fungal pathogens and reduce export rejection rates of Korean grafted cacti. The pathogen specific primer sets F.oF-F.oR, B.CF-B.CR, P.nF-P.nR, and P.cF-P.CR were tested for F. oxysporum, B.cactivora, P. nicotinae, and P. cactorum, respectively. The F.oF-F.oR primer set was designed from the Fusarium ITS region; the B.CF-B.CR and P.nF-P.nR primers respectively from Bipolaris and Phytophthora ITS1; and the P.cF-P.CR primer set from the Ypt1protein gene region. The quarantine fungal pathogen primer pairs were amplified to the specific number of base pairs in each of the following fungal pathogens: 210-bp (F. oxysporum), 510-bp (B. cactivora), 313-bp (P. nicotinae), and 447-bp (P. cactorum). The detection limit for the mono- and multiplex PCR primer sets was 0.1 ng of template DNA under in vitro conditions. Therefore, each primer set successfully diagnosed contamination of quarantine pathogens in export grafted cacti. Consequently, our methodology is a viable tool to screen contamination of the fungal pathogen in exported grafted cacti.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 2006.04a
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• pp.119-122
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• 2006

In both untreated and conventionally stabilized swine manures antibiotic resistant (AR) microorganisms, Staphylococcus-like and Salmonella-like microorganisms were detected. Also pathogens with MAR phynotype were detected. Presence of such microorganisms suggest high level of pathogen-related health risk to farmers who may be in direct contact with the manure and its conventionally stabilized product In contrast the autothermal thermophilic aerobic digestion (ATAD) treatment have efficiently reduced AR and pathogenicity from the swine manure. When soil was fertilized using swine manure and its stabilized products, despite no detection of MAR-exhibiting pathogen-like microorganisms in fertilized soil, potential pathogen-related health risk could not be ruled out from the fertilized soil since the organic fertilization led to increase in AR and pathogenicity in the soil microbial communities. As conclusion, this microbiological study demonstrated that an ATAD process is applicable in control of pathogen-related health risk in livestock manure.

• Journal of Marine Bioscience and Biotechnology
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• v.6 no.1
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• pp.1-7
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• 2014

Marine-derived pathogens threat health and life of human and animals. Therefore, rapid and specific detection methods need to be developed. Here, we summarized various groups of detection methods, including conventional method, flow cytometry, nucleic acid-based method, and protein-based method. In addition, perspective of detection technique was discussed as a unified detection system for pathogens.

• Proceedings of the Microbiological Society of Korea Conference
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• 2003.05a
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• pp.73-76
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• 2003

• 축산식품과학과 산업
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• v.5 no.2
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• pp.50-57
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• 2016