• Title/Summary/Keyword: infectious pathogen

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Development of Enrichment Semi-nested PCR for Clostridium botulinum types A, B, E, and F and Its Application to Korean Environmental Samples

  • Shin, Na-Ri;Yoon, So-Yeon;Shin, Ji-Hun;Kim, Yun Jeong;Rhie, Gi-eun;Kim, Bong Su;Seong, Won Keun;Oh, Hee-Bok
    • 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.

Determination of Neurotoxin Gene Expression in Clostridium botulinum Type A by Quantitative RT-PCR

  • Shin, Na-Ri;Shin, Ji-Hun;Chun, Jeong Hoon;Yoon, So-Yeon;Kim, Bong Su;Oh, Hee-Bok;Rhie, Gi-eun
    • Molecules and Cells
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    • v.22 no.3
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    • pp.336-342
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    • 2006
  • Real time reverse transcription (RT)-PCR was used to quantify the expression of the botulinum neurotoxin type A (BoNT/A) gene (cntA) by normalization with the expression of 16S rRNA. The method were confirmed by monitoring the mRNA levels of cntA during growth in five type A strains. In all but one of the strains the expression of cntA mRNA was maximal in the late exponential phase, and approximately 35-fold greater than in the early exponential phase. The concentration of the extracellular BoNT/A complex detected by ELISA was highest in stationary phase. Sodium nitrite and sorbic acid completely inhibited growth at 20 ppm and $4mg\;ml^{-1}$, respectively. CntA expression became lower in proportion to the concentration of sorbic acid, and this reduction was confirmed by mouse bioassay. Our results show that real time RT-PCR can be used to quantify levels of C. botulinum type A neurotoxin transcripts and to assess the effects of food additives on botulinal risk.

Future Perspectives on New Approaches in Pathogen Detection

  • Li, Peng;Ho, Bow;Ding, Jeak Ling
    • Biomedical Science Letters
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    • v.21 no.4
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    • pp.165-171
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    • 2015
  • Microbial pathogens are responsible for most of the rapidly-spreading deadly infectious diseases against humans. Thus, there is an urgent need for efficient and rapid detection methods for infectious microorganisms. The detection methods should not only be targeted and specific, but they have to be encompassing of potential changes of the pathogen as it evolves and mutates quickly during an epidemic or pandemic. The existing diagnostics such as the antibody-based ELISA immunoassay and PCR methods are too selective and narrowly focused; they are insufficient to capture newly evolved mutant strains of the pathogen. Here, we introduce a fresh perspective on some new technologies, including aptamers and next generation sequencing for pathogen detection. These technologies are not in their infancy; they are reasonably mature and ready, and they hold great promise for unparalleled applications in pathogen detection.

Biological Infectious Watermarking Model for Video Copyright Protection

  • Jang, Bong-Joo;Lee, Suk-Hwan;Lim, SangHun;Kwon, Ki-Ryong
    • Journal of Information Processing Systems
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    • v.11 no.2
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    • pp.280-294
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    • 2015
  • This paper presents the infectious watermarking model (IWM) for the protection of video contents that are based on biological virus modeling by the infectious route and procedure. Our infectious watermarking is designed as a new paradigm protection for video contents, regarding the hidden watermark for video protection as an infectious virus, video content as host, and codec as contagion medium. We used pathogen, mutant, and contagion as the infectious watermark and defined the techniques of infectious watermark generation and authentication, kernel-based infectious watermarking, and content-based infectious watermarking. We experimented with our watermarking model by using existing watermarking methods as kernel-based infectious watermarking and content-based infectious watermarking medium, and verified the practical applications of our model based on these experiments.

The Poly-γ-ᴅ-Glutamic Acid Capsule of Bacillus licheniformis, a Surrogate of Bacillus anthracis Capsule Induces Interferon-Gamma Production in NK Cells through Interactions with Macrophages

  • Lee, Hae-Ri;Jeon, Jun Ho;Rhie, Gi-Eun
    • Journal of Microbiology and Biotechnology
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    • v.27 no.5
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    • pp.1032-1037
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    • 2017
  • The poly-${\gamma}$-$\small{D}$-glutamic acid (PGA) capsule, a major virulence factor of Bacillus anthracis, provides protection of the bacterium from phagocytosis and allows its unimpeded growth in the host. We investigated crosstalk between murine natural killer (NK) cells and macrophages stimulated with the PGA capsule of Bacillus licheniformis, a surrogate of the B. anthracis capsule. PGA induced interferon-gamma production from NK cells cultured with macrophages. This effect was dependent on macrophage-derived IL-12 and cell-cell contact interaction with macrophages through NK cell receptor NKG2D and its ligand RAE-1. The results showed that PGA could enhance NK cell activation by inducing IL-12 production in macrophages and a contact-dependent crosstalk with macrophages.

Performance of the xTAG$^{(R)}$ Gastrointestinal Pathogen Panel, a Multiplex Molecular Assay for Simultaneous Detection of Bacterial, Viral, and Parasitic Causes of Infectious Gastroenteritis

  • Claas, Eric C.;Burnham, Carey-Ann D.;Mazzulli, Tony;Templeton, Kate;Topin, Francois
    • 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.

Construction of Luminescence- and Fluorescence-Tagged Burkholderia pseudomallei for Pathogen Tracking in a Mouse Model

  • Shin, Yong-Woo;Park, Deok Bum;Choi, Myung-Min;Chun, Jeong-Hoon;Seong, Baik-Lin;Rhie, Gi-Eun
    • Journal of Microbiology and Biotechnology
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    • v.28 no.3
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    • pp.498-502
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    • 2018
  • Molecular imaging is a powerful method for tracking various infectious disease-causing pathogens in host organisms. Currently, a dual molecular imaging method that can provide temporal and spatial information on infected hosts at the organism, organ, tissue, and cellular levels simultaneously has not been reported for Burkholderia pseudomallei, a high-risk pathogen that causes melioidosis. In this study, we have established an experimental method that provides spatiotemporal information on infected hosts using luminescent and fluorescent dual-labeled B. pseudomallei. Using this method, we visualized B. pseudomallei infection at the organism, organ, and tissue levels in a BALB/c mouse model by detecting its luminescence and fluorescence. The infection of B. pseudomallei at the cellular level was also visualized by its emitted fluorescence in infected macrophage cells. This method could be an extremely useful and applicable tool to study the pathogenesis of B. pseudomallei-related infectious diseases.

A Marine Bacterium with Animal-Pathogen-Like Type III Secretion Elicits the Nonhost Hypersensitive Response in a Land Plant

  • Boyoung Lee;Jeong-Im Lee;Soon-Kyeong Kwon;Choong-Min Ryu;Jihyun F. Kim
    • The Plant Pathology Journal
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    • v.39 no.6
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    • pp.584-591
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    • 2023
  • Active plant immune response involving programmed cell death called the hypersensitive response (HR) is elicited by microbial effectors delivered through the type III secretion system (T3SS). The marine bacterium Hahella chejuensis contains two T3SSs that are similar to those of animal pathogens, but it was able to elicit HR-like cell death in the land plant Nicotiana benthamiana. The cell death was comparable with the transcriptional patterns of H. chejuensis T3SS-1 genes, was mediated by SGT1, a general regulator of plant resistance, and was suppressed by AvrPto1, a type III-secreted effector of a plant pathogen that inhibits HR. Thus, type III-secreted effectors of a marine bacterium are capable of inducing the nonhost HR in a land plant it has never encountered before. This suggests that plants may have evolved to cope with a potential threat posed by alien pathogen effectors. Our work documents an exceptional case of nonhost HR and provides an expanded perspective for studying plant nonhost resistance.

Creating a Workplace Handbook of Biologically Hazardous Agents (사업장의 생물학적 유해인자 편람 작성 연구)

  • Rim, Kyung-Taek
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.24 no.1
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    • pp.14-37
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    • 2014
  • Objectives: Since information on biological factors in the workplace are currently lacking, I wanted to create a handbook of these factors that would be viewable at a glance as a means to more effectively prevent occupationally-infected diseases. Proper information on biological hazards in the workplace allowing the appropriate recognition of the harmful factors is desperately needed. Methods and Results: In this study, I intended to create a high-utility handbook of biologically hazardous agents in the workplace. To ensure its effectiveness, information and references about biologically hazardous agents in the workplace were analyzed and classified and pathogen safety data sheets(PSDS) sourced from the Public Health Agency of Canada were included. I intended to make it accessible from the point of view of workers and their employers. A more effective classification system of occupational infectious diseases is presented, and biologically hazardous agents were classified according to occupations, industries, infectious diseases, and so on. The handbook consists of 60 major kinds of biologically infectious occupational factors that are expected to be generated in workplaces in Korea, and are focused on practical utility. The pathogen safety data sheets(PSDS) of 192 species were also included. To allow more effective management, domestic and foreign laws and regulations are presented. Conclusions: This case report presents general information on the history and contents of the handbook and PSDS, it will also be useful in workplaces if download from the homepage of OSHRI, KOSHA(oshri.kosha.or.kr/bridge?menuID=901).