• 한국동물위생학회지
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• 제39권4호
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• pp.247-251
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• 2016

Salmonella is one of the most common bacteria that causes heavy losses in swine industry and major causative pathogen of food poisoning in public health. Various methods for the identification of Salmonella such as Gram staining, agglutination test, enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR) have been used. Several studies have demonstrated that Matrix Assisted Laser Desorption Ionization Time of Flight (MALDI TOF) Mass Spectrometry (MS) identification is an efficient and inexpensive method for the rapid and routine identification of isolated bacteria. In this study, MALDI TOF MS could provide rapid, accurate identification of Salmonella spp. from swine compared with end point PCR and real time PCR.

• The Plant Pathology Journal
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• 제22권2호
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• pp.155-160
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• 2006

A strain of Pepper mottle virus (PepMov) was isolated from chili pepper plants in Korea. In host range study, this virus, designated PepMoV-SNU1, shared most characteristics with PepMoV isolates reported previously. Thermal inactivation point ($45^{\circ}C\;to\;75^{\circ}C$) and dilution end point ($10^{-1}\;to\;10^{-4}$) of PepMoV-SNU1 showed differences depending on the propagation hosts. Cylindrical and pinwheel-shaped inclusions were always observed in pepper leaf tissues infected with the virus alone. Unexpectedly, a special structure of pinwheel shaped inclusion surrounded with unknown small spots was also observed in the leaf section when co-infected with a strain of pepper mild mottle virus. The partial sequence of coat protein gene and 3' untranslated region of PepMoV-SNU1 showed 98% identity with those of other PepMoV isolates. A primer pair derived from 3' end of the coat protein gene and poly A tail regions were designed. Optimal detection condition of PepMoV-SNU1 by RT-PCR was tested to determine appropriate annealing temperature and additional volumes of oligo-dT (18-mer), dNTP, and Taq polymerase. Under the optimized condition, an expected 500 Up PCR-product was detected in pepper leaves infected with PepMoV-SNU1 but not in healthy plants.

• 식물병연구
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• 제7권1호
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• pp.1-7
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• 2001

1998년 수원 근교에서 채집한 전형적인 모자이크 병징을 나타내는 수국(Hydragea macrophylla for. otaksa)으로부터 CMV를 분리하고, Hm-CMV라 명명하였다. 기주실험, 물리적 실험, 혈청학적 성질, RNA와 coat protein의 성질, RT-PCR 및 RAP-PCR 분석을 통하여 Hm-CMV의 특성을 분석하였다. 12종의 CMV 지표식물에서 실시한 기주반응실험의 결과, 지금까지 보고된 CMV 계통들의 반응과 특징적인 차이는 인정되지 않았다. Hm-CMV의 물리적 성질은 내열성에서 6$0^{\circ}C$를 보여 기존 CMV들 보다 낮았다. 혈청학적으로 Hm-CMV는 Y-CMV와 융합하는 subgroup I CMV로 분석되었다. SDS-PAGE로부터에 Hm-CMV의 외피단백질은 28 kDa의 band가 확인되었으며, 4종의 게놈 RNA는 Y-CMV와 같은 분자량을 나타냈으나, 위성 RNA는 존재하지 않았다. 수국의 이병엽에서 분리한 dsRNA의 분석 결과도 Y-CMV와 같은 패턴을 보였다. Hm-CMV의 외피단백질유전자에 대한 RT-PCR 분석 결과, 예상된 분자크기의 DNA 증폭이 인정되었으며, PCR 산물을 이용한 EcoR I 및 Msp I을 처리한 결과는 subgroup I CMV의 특성을 나타냈다. 그런, RAP-PCR의 결과, Hm-CMV는 subgroup I내의 다른 계통들과 구분되었다.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2013년도 제45회 하계 정기학술대회 초록집
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• pp.273-273
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• 2013

Recently, Point-of-care (POC) testing microdevices enable to do the patient monitoring, drug screening, pathogen detection in the outside of hospital. Immunochromatographic strip (ICS) is one of the diagnostic technologies which are widely applied to POC detection. Relatively low cost, simplicity to use, easy interpretations of the diagnostic results and high stability under any circumstances are representative advantages of POC diagnosis. It would provide colorimetric results more conveniently, if the genetic analysis microsystem incorporates the ICS as a detector part. In this work, we develop a reverse transcriptase-polymerase chain reaction (RT-PCR) microfluidic device integrated with a ROSGENE strip for colorimetric influenza H1N1 virus detection. The integrated RT-PCR- ROSGENE device is consist of four functional units which are a pneumatic micropump for sample loading, 2 ${\mu}L$ volume RT-PCR chamber for target gene amplification, a resistance temperature detector (RTD) electrode for temperature control, and a ROSGENE strip for target gene detection. The device was fabricated by combining four layers: First wafer is for RTD microfabrication, the second wafer is for PCR chamber at the bottom and micropump channel on the top, the third is the monolithic PDMS, and the fourth is the manifold for micropump operation. The RT-PCR was performed with subtype specific forward and reverse primers which were labeled with Texas-red, serving as a fluorescent hapten. A biotin-dUTP was used to insert biotin moieties in the PCR amplicons, during the RT-PCR. The RT-PCR amplicons were loaded in the sample application area, and they were conjugated with Au NP-labeled hapten-antibody. The test band embedded with streptavidins captures the biotin labeled amplicons and we can see violet colorimetric signals if the target gene was amplified with the control line. The off-chip RT-PCR amplicons of the influenza H1N1 virus were analyzed with a ROSGENE strip in comparison with an agarose gel electrophoresis. The intensities of test line was proportional to the template quantity and the detection sensitivity of the strip was better than that of the agarose gel. The test band of the ROSGENE strip could be observed with only 10 copies of a RNA template by the naked eyes. For the on-chip RT-PCR-ROSGENE experiments, a RT-PCR cocktail was injected into the chamber from the inlet reservoir to the waste outlet by the micro-pump actuation. After filling without bubbles inside the chamber, a RT-PCR thermal cycling was executed for 2 hours with all the microvalves closed to isolate the PCR chamber. After thermal cycling, the RT-PCR product was delivered to the attached ROSGENE strip through the outlet reservoir. After dropping 40 ${\mu}L$ of an eluant buffer at the end of the strip, the violet test line was detected as a H1N1 virus indicator, while the negative experiment only revealed a control line and while the positive experiment a control and a test line was appeared.

• 식물병연구
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• 제12권2호
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• pp.139-143
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• 2006

VC/RT-PCR은 바이러스의 단백질과 PCR 튜브의 재질적 특성을 이용하여 바이러스 이병주의 즙액으로부터 핵산을 지니고 있는 바이러스를 polypropylene에 부착시키고 그 외 PCR 활성을 저해하는 식물 즙액은 PBST로 제거해 주므로써 정확하고 깨끗한 바이러스 진단이 가능하다. 이 방법은 바이러스에 대한 항혈청이 전혀 필요 없으며 포장 시료를 직접 이용해 30 분 이내에 바이러스 핵산을 획득할 수 있어 바이러스 진단이 빠르고 간편하며 경제적이고 감도 높은 실용적인 방법이다. TSWV 에 대한 VC/RT-PCR을 위해 다양한 마쇄 완충액을 시험한 결과 0.5%의 Sodium sulphite를 포함한 0.01M Potassium phosphate (pH 7.0)가 가장 안정적이었으며 VC/RT-PCR을 이용하여 TSWV에 대한 최적의 처리 과정을 확립하였다. TSWV에 최적인 완충액에 마쇄한 바이러스 감염 잎의 조즙액을 이용한 한계 희석 농도는 $10^{-5}$으로 높은 감도를 보여 낮은 농도의 바이러스를 보유한 기주로부터 정확하 게 바이러스를 감지해 낼 수 있게 되었다. 두가지 이상의 바이러스에 감염된 기주로부터 두 가지 이상의 프라이머를 이용해 바이러스를 감지해 내는 다중 진단을 위한 실험 중 식물체의 종류와 바이러스의 Primer의 종류가 진단결과의 정확도에 큰 영향을 미치며 이러한 결과는 식물체 즙액을 바로 이용하는 VC/RT-PCR 방법은 물론 식물체로부터 Total RNA를 따로 분리하여 RT-PCR을 이용한 결과에서도 동일하게 나타났다. 따라서 편리하고 실용 적인 VC/RT-PCR법의 정확성을 최대화시키기 위해서는 다중 진단을 저해하는 식물체적 원인과 Primer 사이의 간섭 현상에 대한 연구가 더 진행될 것이며 이러한 원인을 충분히 제거해 줄 수 있는 마쇄 완충액을 개발하고 프라이머 제작을 더욱 신중히 해야 할 것이다.

• The Plant Pathology Journal
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• 제26권4호
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• pp.328-339
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• 2010

A tobamovirus, Ribgrass mosaic virus (RMV), was identified newly from chinese cabbage (Brassica campestris L. pekinensis) in Korea. Virus disease incidence of RMV on chinese cabbage was 37.9% in alpine area on August in 1993. RMV induced the symptoms of necrotic ring spots, necrotic streak on midrib and malformation. RMV, Ca1 and Ca3 isolate, could infect 35 species out of 45 plants including Chenopodium amaranticolor. Physical properties of RMV Ca1 isolate were very stable as 10.8 over for dilution end point, $95^{\circ}C$ for temperature inactivation point and 18 weeks for longevity in vitro. RMV had the soil transmission rate of 75.0% for the chinese cabbages, 'Chunhawang' and 'Seoul' cultivars. The purified virions of RMV had the typical ultraviolet absorption spectrum of maximum at 260 nm and minimum at 247 nm. RMV of Ca1 isolate was related serologically with antisera of Tobacco mosaic virus (TMV)-Cym, TMV-O and Pepper mottle virus, but not related with antiserum of Odontoglossum ring spot virus. coat protein gene of RMV-Ca1, sized 473 nucleotides, encoded 158 amino acid residues. Nucleotide identity of RMV-Ca1 CP gene was 96.4% with RMV-Shanghai (GenBank accession No. of AF185272) from China and 96.0% with RMV-Impatiens (GenBank accession No. of AM040974) from Germany. Identity of amino acids between RMV-Ca1 and the two RMV isolates was 96.8%. Specific three primers were selected for rapid and easy genetic detection of RMV using Virion Captured (VC)/RT-PCR method.

• Journal of Microbiology and Biotechnology
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• 제21권6호
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• pp.659-664
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• 2011

Staphylococcus aureus (SA) bacteremia is associated with high mortality, and often results in metastatic infections. The methicillin-resistant SA (MRSA) is an urgent health care issue, as nosocomial infections with these bacteria represent limited treatment alternatives. Samples of whole blood containing challenge inoculums of SA and MRSA strains were passed through columns packed with surfaceheparinized polyethylene beads. The bound bacteria were eluted and quantitatively determined by culturing and by real-time PCR. Significant amounts of both SA and MRSA adhered to the heparinized beads (more than 65% of inoculated bacteria). After rinsing with buffer at high ionic strength, viable bacteria or bacterial DNA were eluted from the columns, indicating that the binding was specific. The conclusions that can be made from these experiments are that, as earlier reported in the literature, the high affinity of SA to heparin is retained in whole blood, and MRSA in whole blood binds to heparin with similar or higher affinity than SA. It should be possible to lower the amount of SA and/or MRSA from the blood of infected patients to levels that could be taken care of by the immune system. In previous studies, we have shown that passing blood from septic patients over beads coated with end-point-attached, biologically active heparin is a useful technique for regulating the levels of heparinbinding cytokine. These findings in combination with the present findings indicate the possibility of creating an apheresis technology for treatment of sepsis caused by SA and/or MRSA.

• 생물정신의학
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• 제8권1호
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• pp.106-110
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• 2001

Background : Dopamine receptors have been regarded as a strong candidate involved in etiology of schizophrenia and a target for various antipsychotic drugs. The purpose of our study was to investigate whether dopamine $D_1$ receptor(DRD1) gene polymorphisms would predict the treatment response to antipsychotics in schizophrenia. Method : One hundred thirty-four schizophrenic patients, who met DSM-IV criteria for schizophrenia were entered into a 48 -week study. The psychopathology of the patients was assessed at baseline, 12th, 24th 48th weeks of treatment by PANSS. Responders were defined by a 20% of the reduction in total PANSS score at end point. The genomic DNA fragment corresponding to nucleotides of dopamine $D_1$ receptor gene was amplified by polymerase chain reaction(PCR). Result: Neither allelic frequencies nor genotypes for dopamine $D_1$ receptor differed significantly between responders and non-responders. Also, there was no difference of changes of PANSS scores among three genotype groups of the dopamine $D_1$ receptor. Conclusion : Allelic variation in the dopamine $D_1$ gene is not associated with individual differences in antipsychotic response.

• Asian Pacific Journal of Cancer Prevention
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• 제13권12호
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• pp.6155-6157
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• 2012

Background: Mutations in the MAPK (Mitogen Activated Protein Kinase) signaling pathway - EGFR/Ras/RAF/MEK have been associated with the development of several carcinomas. ERK2, a downstream target of the MAPK pathway and a founding member of the MAPK family is activated by cellular signals emanating at the cell membrane. Activated ERK2 translocates into the nucleus to transactivate genes that promote cell proliferation. MKP - a dual specific phosphatase - interacts with activated ERK2 via the common docking (CD) domain of the later to inactivate (dephosphorylate) and effectively terminate further cell proliferation. A constitutively active form of ERK2 carrying a single point mutation - E322K in its CD domain, was earlier reported by our laboratory. In the present study, we investigated the prevalence of this CD domain E322K mutation in 88 well differentiated OSCC tissue samples. Materials and Method: Genomic DNA specimens isolated from 88 oral squamous cell carcinoma tissue samples were amplified with primers flanking the CD domain of the ERK2 gene. Subsequently, PCR amplicons were gel purified and subjected to direct sequencing to screen for mutations. Results: Direct sequencing of eighty eight OSCC samples identified an E322K CD domain mutation in only one (1.1%) OSCC sample. Conclusions: Our result indicates that mutation in the CD domain of ERK2 is rare in OSCC patients, which suggests the role of genetic alterations in other mitogenic genes in the development of carcinoma in the rest of the patients. Nevertheless, the finding is clinically significant, as the relatively rare prevalence of the E322K mutation in OSCC suggests that ERK2, being a common end point signal in the multi-hierarchical mitogen activated signaling pathway may be explored as a viable drug target in the treatment of OSCC.

• 한국지하수토양환경학회지:지하수토양환경
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• 제10권2호
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• pp.52-58
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• 2005

본 연구는 사염화에틸렌(Perchloroethylene) 또는 트리클로로에틸렌(Trichloroethylene)으로 오염된 국내 지하수 내에 국외에서 보고된 환원성 탈염소화 미생물의 존재 유무와 사염화에틸렌의 생물학적 탈염소화 활성도를 평가하였다. 마이크로코즘 테스트(microcosm tests)는 4개의 오염지역(창원 A, 창원 B, 부천 및 양산) 지하수와 다양한 전자공여체 (sodium lactate, sodium propionate, sodium butyrate, sodium fumarate)를 이용하여 수행하였다. 단일 전자공여체 와 창원 A 혹은 창원 B 지하수를 주입한 전 마이크로코즘에서 사염화에틸렌 완전 탈염소화 분해 시 발생하는 최종 산물인 에틸렌이 배양 90일 후에 검출되었고, 부천 혹은 양산 지역의 지하수를 주입한 마이크로코즘에서는 배양 90 일 후 시스-1,2-디클로로에틸렌(cis-1 ,2-Dichloroethylene)만이 검출되었고, 염화비닐(Vinyl chloride) 과 에틸렌은 검출되지 않았다. 완전 탈염소화 생분해가 확인된 창원 B 지역 지하수와 불완전 탈염소화 생분해가 확인된 양산 지역 지하수 내 미생물 군집을 비교하기 위해 분자생물학적 방법을 이용한 실험을 수행하였다. 창원 B 지역 지하수의 클론 라이브러리(Clone library)에서 사염화에틸렌 완전 탈염소화 미생물, uncultured bacterium clone DCE47과 매우 유사한 염기서열 클론이 확인되었다. 그러나 양산 지역의 클론 라이브러리에서는 기존의 염화에틸렌 탈염소화 미생물과 유사한 염기서열 클론이 확인되지 않았다. 본 연구 결과를 통하여 국내 일부 지역의 지하수 내에 사염화에틸렌을 완전 탈염소화하여 무해한 에틸렌으로 분해하는 미생물이 존재함을 확인하였고, 적절한 전자공여체를 공급하는 경우 그 분해 활성도가 증가함을 확인하였다. 이 결과는 사염화에틸렌 혹은 트리클로로에틸렌으로 오염된 국내 지하수를 경제적인 공법인 환원성 탈염소화 생물학적 공정으로 복원할 수 있는 기능성을 보여주는 중요한 지표라고 사료된다.