• 대한수의학회지
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• 제52권2호
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• pp.75-82
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• 2012

We have developed and optimized a multiplex polymerase chain reaction (mPCR) for simultaneous detection of Brucella, Salmonella and Leptospira with high sensitivity and specificity. Three pairs of oligonucleotide primers were designed to specifically amplify the targeted genes of Salmonella, Leptospira and Brucella species with sizes of 521, 408 and 223 bp, respectively. The mPCR did not produce any nonspecific amplification products when tested against 15 related species of bacteria. The sensitivity of the mPCR was 100 fg for Brucella and 1 pg for both Salmonella and Leptospira species. In the field application, kidney, liver and spleen were collected from wild rats and stray cats and examined by mPCR. The high specificity and sensitivity of this mPCR assay provide a valuable tool for diagnosis and for the simultaneous and rapid detection of three zoonotic bacteria that cause disease in both humans and animals. Therefore, this assay could be a useful alternative to the conventional method of culture and single PCR for the detection of each pathogen.

• Journal of Microbiology and Biotechnology
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• 제29권8호
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• pp.1184-1192
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• 2019

The influenza A virus is a highly infectious respiratory pathogen that sickens many people with respiratory disease annually. To prevent outbreaks of this viral infection, an understanding of the characteristics of virus-host interaction and development of an anti-viral agent is urgently needed. The influenza A virus can infect mammalian species including humans, pigs, horses and seals. Furthermore, this virus can switch hosts and form a novel lineage. This so-called zoonotic infection provides an opportunity for virus adaptation to the new host and leads to pandemics. Most influenza A viruses express proteins that antagonize the antiviral defense of the host cell. The non-structural protein 1 (NS1) of the influenza A virus is the most important viral regulatory factor controlling cellular processes to modulate host cell gene expression and double-stranded RNA (dsRNA)-mediated antiviral response. This review focuses on the influenza A virus NS1 protein and outlines current issues including the life cycle of the influenza A virus, structural characterization of the influenza A virus NS1, interaction between NS1 and host immune response factor, and design of inhibitors resistant to the influenza A virus.

• Journal of Microbiology and Biotechnology
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• 제29권10호
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• pp.1543-1552
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• 2019

Salmonella is a common zoonotic and foodborne pathogen that causes high morbidity and mortality in developing countries. In this study, we established and validated a polymerase spiral reaction (PSR) assay which targeted the conserved invasion gene (invA) of Salmonella by SYBR Green I indicator methods. Subsequently, assays for determination of the optimal conditions for optimal specificity and sensitivity of PSR were performed. We performed comprehensive evaluations using loop-mediated isothermal amplification (LAMP) and real-time PCR. A total number of 532 samples of daily food were analyzed by PSR. Twenty-seven bacterial strains were tested in the specificity assay, from which positive results were obtained only for 14-Salmonella strains. However, none of the 13 non-Salmonella strains was amplified. Similarly with LAMP and real-time PCR, the detection limit of the PSR assay was 50 CFU/ml. The PSR method was also successfully applied to evaluate the contamination with Salmonella in 532 samples of daily food, corroborating traditional culture method data. The novel PSR method is simple, sensitive, and rapid and provides new insights into the prevention and detection of foodborne diseases.

• Journal of Veterinary Science
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• 제23권2호
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• pp.28.1-28.18
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• 2022

Enteropathogenic Escherichia coli (EPEC) is a major cause of infantile diarrhea in developing countries. However, sporadic outbreaks caused by this microorganism in developed countries are frequently reported recently. As an important zoonotic pathogen, EPEC is being monitored annually in several countries. Hallmark of EPEC infection is formation of attaching and effacing (A/E) lesions on the small intestine. To establish A/E lesions during a gastrointestinal tract (GIT) infeciton, EPEC must thrive in diverse GIT environments. A variety of stress responses by EPEC have been reported. These responses play significant roles in helping E. coli pass through GIT environments and establishing E. coli infection. Stringent response is one of those responses. It is mediated by guanosine tetraphosphate. Interestingly, previous studies have demonstrated that stringent response is a universal virulence regulatory mechanism present in many bacterial pathogens including EPEC. However, biological signficance of a bacterial stringent response in both EPEC and its interaction with the host during a GIT infection is unclear. It needs to be elucidated to broaden our insight to EPEC pathogenesis. In this review, diverse responses, including stringent response, of EPEC during a GIT infection are discussed to provide a new insight into EPEC pathophysiology in the GIT.

• Parasites, Hosts and Diseases
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• 제62권1호
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• pp.151-156
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• 2024

Bovine borreliosis, caused by Borrelia theileri which is transmitted via hard tick bites, is associated with mild clinical symptoms, such as fever, lethargy, hemoglobinuria, anorexia, and anemia. Borrelia theileri infects various animals, such as cattle, deer, horses, goats, sheep, and wild ruminants, in Africa, Australia, and South America. Notably, no case of B. theileri infection has been reported in Korean cattle to date. In this study, 101 blood samples were collected from a Korean indigenous cattle breed, among which 1.98% tested positive for B. theileri via nested PCR. The obtained sequences exhibited high homology with B. theileri strains identified in other regions. Phylogenetic analysis of 16S rRNA confirmed the B. theileri group affiliation; however, flagellin B sequences exhibited divergence, potentially due to regional evolutionary differences. This study provides the first molecular confirmation of B. theileri infection in Korean livestock. Further isolation and nucleotide sequence analyses are necessary to better understand the presence of B. theileri strains in cows in Korea.

• Journal of Microbiology and Biotechnology
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• 제18권3호
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• pp.585-590
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• 2008

Human tropic Porcine Endogenous Retroviruses (PERVs) are the major concern in zoonosis for xenotransplantation because PERVs cannot be eliminated by specific pathogen-free breeding. Recently, a PERV A/C recombinant with PERV-C bearing PERV-A gp70 showed a higher infectivity (approximately 500-fold) to human cells than PERV-A. Additionally, the chance of recombination between PERVs and HERVs is frequently stated as another risk of xenografting. Overcoming zoonotic barriers in xenotransplantation is more complicated by recombination. To achieve successful xenotransplantation, studies on the recombination in PERVs are important. Here, we cloned and sequenced proviral PERV env sequences from pig gDNAs to analyze natural recombination. The envelope is the most important element in retroviruses as a pivotal determinant of host tropisms. As a result, a total of 164 PERV envelope genes were cloned from pigs (four conventional pigs and two miniature pigs). Distribution analysis and recombination analysis of PERVs were performed. Among them, five A/B recombinant clones were identified. Based on our analysis, we determined the minimum natural recombination frequency among PERVs to be 3%. Although a functional recombinant envelope clone was not found, our data evidently show that the recombination event among PERVs may occur naturally in pigs with a rather high possibility.

• 생명과학회지
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• 제30권9호
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• pp.749-762
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• 2020

돼지인플루엔자는 동물에서 사람에게 감염할 수 있는 인수공통전염병이다. 우리는 2019년 한국 돼지농장에서 호흡기 증상을 보이는 돼지에서 3주의 H1N2형 인플루엔자바이러스를 분리하였다. 유전자 분석결과, 이들 바이러스의 8개 유전자 중 PA 및 NP 유전자는 2009 대유행 H1N1 인플루엔자 유래였고, 나머지 유전자는 돼지에 유행하는 H3N2 및 H1N2 인플루엔자 유래 유전자를 가진 재조합 바이러스 이었다. 분리된 H1N2 바이러스를 마우스에 접종한 결과, 마우스는 17% 정도 체중이 감소하였고, 염증 세포들이 침윤한 간질성 폐렴 증상을 보였다.

• 한국식품위생안전성학회지
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• 제28권2호
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• pp.95-98
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• 2013

Streptococcus suis (S. suis) is a major swine pathogen and an emerging zoonotic agent and is an increasing public health problem across Asia. The present study was undertaken to estimate the antibacterial effect of GR extract and therapeutic effect of GR extract against S. suis infection in mice. At the concentration of GR extract 2.5 mg/ml, the antibacterial effect was not shown on S. suis. However, the antibacterial effect against S. suis was observed at the concentration of GR extract 5.0 mg/ml. Oral administration of GR extract at the dose of 10 mg/kg showed a therapeutic effect for S. suis infected BALB/c mice. The mortality of GR extract-treated mice at the concentration of 5, 10 and 20 mg/kg was 80%, 70%, and 50% at 12 days, respectively, while that of untreated mice was 100% at 8 days after a lethal dose of S. suis infection. The results of our study strongly indicate that GR extract has potential as an effective for S. suis infection in mice.

• IMMUNE NETWORK
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• 제21권1호
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• pp.11.1-11.10
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• 2021

Coronavirus causes an infectious disease in various species and crosses the species barriers leading to the outbreak of zoonotic diseases. Due to the respiratory diseases are mainly caused in humans and viruses are replicated and excreted through the respiratory tract, the nasal fluid and sputum are mainly used for diagnosis. Early diagnosis of coronavirus plays an important role in preventing its spread and is essential for quarantine policies. For rapid decision and prompt triage of infected host, the immunochromatographic assay (ICA) has been widely used for point of care testing. However, when the ICA is applied to an expectorated sputum in which antigens are present, the viscosity of sputum interferes with the migration of the antigens on the test strip. To overcome this limitation, it is necessary to use a mucolytic agent without affecting the antigens. In this study, we combined known mucolytic agents to lower the viscosity of sputum and applied that to alpha and beta coronavirus, porcine epidemic diarrhea virus (PEDV) and Middle East respiratory syndrome coronavirus (MERS-CoV), respectively, spiked in sputum to find optimal pretreatment conditions. The pretreatment method using tris(2-carboxyethyl)phosphine (TCEP) and BSA was suitable for ICA diagnosis of sputum samples spiked with PEDV and MERS-CoV. This sensitive assay for the detection of coronavirus in sputum provides an useful information for the diagnosis of pathogen in low respiratory tract.

• 한국식품위생안전성학회지
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• 제34권6호
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• pp.576-582
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• 2019

Staphylococcus pseudintermedius는 개에서 기회감염을 유발하는 병원체이며, 공중보건학적으로도 주요한 인수공통 병원체이다. 개에서 분리된 S. pseudintermedius 균주들은 주로 항생제 내성 및 개에서 피부 감염을 유발하는 주요 원인균으로 연구되어 왔지만, 가축에서 분리된 S. pseudintermedius 균주들의 항생제 내성 및 장내 독소 생성에 대한 정보는 매우 제한적이다. 본 연구에서는 개, 돼지, 육우에서 분리된 S. pseudintermedius 균주들에서 18가지의 장내 독소 (staphylococcal enterotoxin; SE) 유전자와 toxic shock syndrome toxin 유전자(tst-1)의 분포양상을 조사하였다. 또한, S. pseudintermedius 균주들의 항생제 내성 양상과 더불어 mecA 유전자 및 SCCmec type 또한 확인하였다. 육우에서 분리한 하나의 균주를 제외한 모든 개와 돼지 분리주 들이 4개 이상의 항생제에 내성을 보였으며, 개에서 분리된 6개의 균주 중 4개의 S. pseudintermedius 균주들이 메티실린 내성과 더불어 SCCmec V를 가진 것으로 확인 되었다. 총 11개의 SE 유전자들 (seb, sec, see, seg, sei, sej, sel, seo, sep, seq, seu) 및 tst-1가 개, 돼지 및 육우로부터 분리된 S. pseudintermedius 균주들에서 확인 되었으며, 대부분의 분리주들 (83%)에서 2개 이상의 SE 유전자들이 확인 되었고, 그 중 sel (42%) 및 sep (42%)가 가장 빈번하게 검출 되었다. 본 연구를 통하여 반려견에서 뿐만 아니라 주요 가축에서 존재하는 S. pseudintermedius 균주들에서 높은 항생제 내성 양상을 확인 하였으며, 항생제 내성과 더불어 여러 staphylococcal enterotoxin 및 tst-1 유전자들을 전파할 가능성을 확인 하였다.