• 대한수의학회지
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• 제44권1호
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• pp.83-88
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• 2004

Galectin family, endogenous ${\beta}$-galactoside-binding animal lectins, is known for the role in cell differentiation, morphogenesis, apoptosis and tumorigenesis. Galectin-3, one of family member, has been studied for its role in cell differentiation and tumor metastasis, and for its expression on epithelial cells of colon and mast cells but not in brain. Several reports, however, suggest its expression in brain including as a prion binding protein. In this report we explored possibility of galectin-3 expression in brain tissue. With Western blot and RT-PCR with rat brain tissues, we could detect galectin-3 that was not shown by conventional immunohistochemistry. Our results indicated galectin-3 was expressed in brain, and substantiate the previous report on galecin-3 as a prion-related protein in brain.

• 대한임베디드공학회논문지
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• 제7권1호
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• pp.41-52
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• 2012

Recently, FlexRay was developed to replace controller area network (CAN) protocol in chassis networking systems, to remedy the shortage of transmission capacity and unsatisfactory real-time transmission delay of conventional CAN. FlexRay network systems require correct synchronization and complex scheduling parameters. However, because platform configuration register (PCR) setting and message scheduling is complex and bothersome task, FlexRay is more difficult to implement in applications than CAN protocol. To assist a network designer for implementing FlexRay cluster, this paper presents an analysis of FlexRay platform configuration register and automatic generation program of PCR. To demonstrate the feasibility of the automatic generation program, we evaluated its performance using experimental testbed.

• 한국동물위생학회지
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• 제41권1호
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• pp.29-39
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• 2018

In this study, we developed a sensitive and specific reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid visual detection of foot-and-mouth disease virus (FMDV) circulated in Korea. The RT-LAMP was completed in 40 min at $62^{\circ}C$ and the results of the assay were directly detected by naked eye without any detection process. The assay specifically amplified all 7 serotypes of FMDV RNAs but not amplified other viral and cellular nucleic acids. The sensitivity of the RT-LAMP was $10^2$, $10^3$ and $10^3TCID_{50}/mL$ for serotype O, A and Asia 1 FMDV, respectively, which was comparable to conventional reverse transcription polymerase chain reaction (RT-PCR) and relatively lower than that of real time quantitative RT-PCR (qRT-PCR). Clinical evaluation of the RT-LAMP using different serotypes of Korean and foreign FMDV strains showed a 100% (35/35) agreement with the results of the RT-PCR and qRT-PCR. These results indicated that RT-LAMP assay developed in this study could be a valuable diagnostic method for FMDV monitoring and surveillance.

• Parasites, Hosts and Diseases
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• 제34권1호
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• pp.15-20
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• 1996

이 실아메바(Entamoeba histoIytica)와 동형 아메바(Entamoebc dispar. 동형아메바)의 포낭은 형태학적으로 구분이 안되어 종 감별에 관하여 논란이 있어 왔다. 최근에 이 둘이 별종이며 특히 이질아메바는 병원성이고 동형아메바는 비병윈성입이 확인퇴어 그 감별이 중요한 의미를 갖게 피었나 이 연구에서는 우리 나라의 아메바 분리체를 중합효소 반응과 제한효소 반응을 이용하여 두 종으로 감별하였다. 1994-1995년에 대변을 통강의 방법으로 검색하여 포낭이나 영양형이 발견된 검체를 로빈슨 배지에서 배양하고 배양된 영양형에서 DNA를 분리하였다. PI 유전자 염기서열 중에서 시발페(primer)를 만득어 중합효소 반응으로 482 bp 크기의 산물을 얹고 이를 제한효소(Tuq I, Xmn I, Acc I)로 처리하였다. 또한 Xmn I과 Acc I 제한 효소의 특이 염기서열온 고함하는 시발체를 제작하여 따로 중합효소 반응을 시행하였다. 그 결과 13개 분리체 중에서 S9, S12, YS-6, YS-27의 482 bp 산물은 Taq I과 Xmn I 의하여 그 외의 분리체 산물은 Acc I에 의하여 절단되었다. 이 결과는 특이 염기서열 시발체의 중합효소 반응에서 얻은 산물과 일치하였다, 이 결과에 의하여 분리체 S9, S12, YS-6는 대장염 창자에서. YS-27은 간농양 환자에서 분리한 병원성의 이질아메바(E. histolytica)이고 분리체 S1, S3, S11, S15, S16, S17, S20, YS-17, YS-44는 부증상의 포낭배출자에서 얻은 비병원성의 동형아메바(E. nispur)로 구별할 수 있었다. S1은 설사 환자에서 얻은 분리체 이지탄 동형아메바임을 확인하였고 따라서 이 환자의 설사는 다른 원인에 의한 것으로 판단 된다. 이로써 비병원성인 동형아메바가 우리 나라에서도 병원성 이질아메바보다 더 흔하게 존재한다는 것을 처음으로 기록하며 E. dispar의 우리 말 이름을 동형아메바로 제안한다.

• 생태와환경
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• 제54권3호
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• pp.199-208
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• 2021

메타바코딩을 이용한 환경 DNA 분석은 검출 감도가 높아 어류의 생물다양성 평가 및 멸종위기종의 검출에 유용한 기술이다. 이번 연구는 메타바코딩을 이용해 우리나라 담수어류를 대상으로 높은 검출 효율을 보일 수 있는 적합한 분석방법을 확인하기 위해 4가지 분석조건별, 즉 필터(cellulose nitrate filter, glass fiber filter), 추출 키트(DNeasy^® Blood & Tissue Kit, DNeasy^® PowerWater Kit), 프라이머 조합(12S rDNA, 16S rDNA) 그리고 PCR 방법(conventional PCR, touchdown PCR)로 나타나는 Operational Taxonomic Units(OTUs) 수와 종 조성을 비교하였다. Glass fiber filter와 DNeasy^® Tissue & Blood Kit를 이용해 추출한 시료는 12S rDNA와 16S rDNA 프라이머 조합에서 담수어류 OTUs가 가장 많이 검출되었다. 모든 분석조건 중 프라이머 조합에서만 조기어강(Class Actinopterygii) 평균 OTUs 수에서 통계적으로 유의한 차이를 보였고(Non-parametric Wilcoxon Signed Ranks Test, p=0.005), 담수어류 평균 OTUs 수는 유의하지 않았다. 종 조성 비교 결과 역시 프라이머 조합에서 유의한 차이를 보였고(PERMANOVA, Pseudo-F=6.9489, p=0.006), 나머지 분석조건에서는 유의한 차이를 보이지 않았다. NMDS 분석 결과 종 조성은 유사도 65% 기준에서 프라이머 조합에 따라 묶였고, 16S rDNA 프라이머 세트는 주로 멸종위기종인 모래주사(Microphysogobio koreensis), 꼬치동자개(Pseudogobio brevicorpus)가 기여하였고, 12S rDNA 프라이머 세트는 주로 일반종인 피라미(Zacco platypus), 꺽지(Coreoperca herzi) 등이 기여한 것으로 나타났다. 본 연구는 국내 하천에서 채취한 시료에 대한 메타바코딩을 이용한 종 다양성 분석의 기초정보를 제공한다.

• The Plant Pathology Journal
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• 제31권3호
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• pp.219-225
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• 2015

The primary step for efficient control of viral diseases is the development of simple, rapid, and sensitive virus detection. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) has been used to detect viral RNA molecules because of its simplicity and high sensitivity for a number of viruses. RT-LAMP for the detection of Potato virus X (PVX) was developed and compared with conventional reverse transcription polymerase chain reaction (RT-PCR) to demonstrate its advantages over RT-PCR. RT-LAMP reactions were conducted with or without a set of loop primers since one out of six primers showed PVX specificity. Based on real-time monitoring, RT-LAMP detected PVX around 30 min, compared to 120 min for RT-PCR. By adding a fluorescent reagent during the reaction, the extra step of visualization by gel electrophoresis was not necessary. RT-LAMP was conducted using simple inexpensive instruments and a regular incubator to evaluate whether RNA could be amplified at a constant temperature instead of using an expensive thermal cycler. This study shows the potential of RT-LAMP for the diagnosis of viral diseases and PVX epidemiology because of its simplicity and rapidness compared to RT-PCR.

• 한국환경보건학회지
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• 제42권3호
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• pp.189-195
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• 2016

Objectives: To ensure the microbiological safety of groundwater, it was confirmed whether waterborne pathogenic bacteria in groundwater samples tested positive for total coliforms in the Chungcheongnam-do Province region. Methods: Total colony counts, total coliforms and fecal coliforms were tested according to the process mandated by the drinking water quality testing standards of Korea. DNA was extracted from the samples, tested positive for total coliforms, and then subjected to real-time PCR to detect waterborne pathogenic bacteria. Results: A total of 115 samples were inadequate for drinking water. Thirty-one cases (27%) showed positive for fecal coliforms and nine cases (7.8%) showed total colony counts exceeding drinking water standards. Twenty-seven cases (23.5%) showed three items (total colony counts, total coliforms and fecal coliforms). Using the real-time PCR method, waterborne pathogens were detected in 57 cases (49.6%) in 115 samples. Seventy-eight cases of waterborne pathogenic bacteria were detected (including duplications): 27 cases of pathogenic E. coli (EPEC (19), ETEC (5), EHEC (1), EAEC (1) and EIEC (1)); 45 of Bacillus cereus; two of Yersinia spp.; two of Salmonella spp.; one of Staphylococcus aureus; one of Clostridium perfringens. Conclusion: The real-time PCR method can offer rapid and accurate detection of waterborne pathogenic bacteria. Therefore, this assay could be an alternative to conventional culture methods and can further ensure the microbiological safety of groundwater.

• 대한수의학회지
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• 제57권2호
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• pp.79-88
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• 2017

The study of canine vector-borne diseases in the Philippines started in the 1970s but only gained interest in the past decade. Characterization of such diseases in the Philippines remains incomplete, thus, it is necessary to obtain additional information on the prevalence and diversity of canine tick-borne diseases in the country. In this study, blood samples were obtained at two veterinary clinics in Metro Manila, Philippines from 114 dogs suspected of having canine tick-borne pathogens. Polymerase chain reaction (PCR) was performed on whole blood DNA extracts followed by sequencing, and the following pathogens were detected: Hepatozoon (H.) canis (5.26%), Babesia (B.) vogeli (5.26%), Ehrlichia (E.) canis (4.39%), and Anaplasma platys (3.51%). Additionally, a set of multiplex PCR primers were developed to detect H. canis, Babesia spp. (B. canis and B. vogeli), and E. canis in canine blood. Multiplex and conventional single-reaction PCR results for the 114 dog blood samples were similar, except for one H. canis sample. Multiplex PCR is, therefore, a useful tool in screening infected dogs in veterinary clinics. This study's results, together with those of previous studies in the country, show that canine vector-borne pathogens are an emerging veterinary concern in the Philippines.

• The Plant Pathology Journal
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• 제34권2호
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• pp.104-112
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• 2018

Accurate and rapid detection of bacterial plant pathogen is the first step toward disease management and prevention of pathogen spread. Bacterial plant pathogens Clavibacter michiganensis subsp. nebraskensis (Cmn), Pantoea stewartii subsp. stewartii (Pss), and Rathayibacter tritici (Rt) cause Goss's bacterial wilt and blight of maize, Stewart's wilt of maize and spike blight of wheat and barley, respectively. The bacterial diseases are not globally distributed and not present in Korea. This study adopted comparative genomics approach and aimed to develop specific primer pairs to detect these three bacterial pathogens. Genome comparison among target pathogens and their closely related bacterial species generated 15-20 candidate primer pairs per bacterial pathogen. The primer pairs were assessed by a conventional PCR for specificity against 33 species of Clavibacter, Pantoea, Rathayibacter, Pectobacterium, Curtobacterium. The investigation for specificity and sensitivity of the primer pairs allowed final selection of one or two primer pairs per bacterial pathogens. In our assay condition, a detection limit of Pss and Cmn was $2pg/{\mu}l$ of genomic DNA per PCR reaction, while the detection limit for Rt primers was higher. The selected primers could also detect bacterial cells up to $8.8{\times}10^3cfu$ to $7.84{\times}10^4cfu$ per gram of grain seeds artificially infected with corresponding bacterial pathogens. The primer pairs and PCR assay developed in this study provide an accurate and rapid detection method for three bacterial pathogens of grains, which can be used to investigate bacteria contamination in grain seeds and to ultimately prevent pathogen dissemination over countries.

• 대한수의학회지
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• 제39권2호
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• pp.343-352
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• 1999

Vancomycin resistant enterococci (VRE) have emerged as an important nosocomial pathogen. Since 1989 the Center for Disease Control, United States, has reported a rapid increase in the incidence of enterococcal bacteremia and endocarditis infection by VRE. It was suggested that the use of avoparcin was associated with the appearance of VRE in animal husbandry. To date, several detection methods have been used based on conventional methods of culture and gene detection. However, these methods have some limitations such as time-consuming, laborious and additional differential needs. Therefore, In this study a multiplex PCR method was established to detect and differentiate resistance types of enterococci which specifically amplify the four van genes encoding vancomycin resistance elements. Using the method, we investigated the incidence rates and types of VRE from farms using or not using avoparcin. A total of 1091 animal fecal samples were collected from 70 pig and 32 poultry farms. A total of 425 of enterococci were isolated from samples. Of the 425 isolates, 11 of the them showed a pattern of high-level vancomycin resistance (MIC : $64{\sim}256{\mu}g/ml$) which was associated with the presence of the vanA or vanB gene. Fifty-seven isolates showed a pattern of low-level vancomycin resistance (MIC : $3{\sim}8{\mu}g/ml$) associated with the vanC-1 or vanC-2 gene. Interestingly, all isolates with high-level vancomycin resistance were from farms that have never used avoparcin. Moreover, the high-level VRE isolation rate in Korea (2.58%) was much lower than that of other countries (50% in England, 7% in Belgium) where avoparcin have been used. In conclusion, the multiplex PCR method established in this study could be applied for detection of VRE.