• Title/Summary/Keyword: PCR 동정

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Isolation of Nontuberculous Mycobacteria (NTM) from Air Conditioner Dust (에어컨 먼지내 비결핵 항산균의 동정)

  • Choi, Seung Gu;Choi, Myeong Sik
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.4
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    • pp.435-438
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    • 2017
  • Nontuberculous mycobacteria (NTM) contains M. avium-intracellular complex (MAC), M. fortuitum, M. chelonae, M. abscessus, M. kansasii, etc., which causes infections in swine, poultry, and other animals. These bacteria are opportunistic pathogens that can contaminate people, and in doing so, cause various social economic problems. This study proved that NTM are also present in air conditioners of multiuse facilities in frequently visited populated areas. The results of this study are meaningful because they showed that pathogenic microbes can live in the surroundings and cause diseases to people with impaired immunity. This study used the AFB stain, L-J medium culture, and PCR method for the detection of MTB and NTM in air conditioner dust. MTB was not detected in any of the collected samples, while NTM was detected in 2 out of 40 samples (5%). Most people living in the modern environment cannot avoid being exposed to air conditioners so special attention is needed for sanitary inspections and the management of air conditioners. Moreover, it is important to study NTM through various individuals, accumulate the related data and establish the methods and standards of hygiene management system to reduce NTM infections in public areas. Moreover, it would be necessary to identify the precise species of NTM through DNA sequencing and evaluate the effects of NTM on the pulmonary tuberculosis in immunodeficiency patients.

Comparison of Harboring the Resistance Gene and Disc Diffusion Susceptibility Test Result in Staphylococcus pseudintermedius from the Bacterial Dermatitis (세균성 피부염 개에서 분리된 Staphylococcus pseudintermedius에서 항생제 감수성 검사와 내성 유전자 획득의 비교)

  • Jang, Hye-Jin;Son, Hyoung-Won;Kang, Hyo-Min;Han, Jae-Ik;Na, Ki-Jeong
    • Journal of Veterinary Clinics
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    • v.32 no.2
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    • pp.158-161
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    • 2015
  • Bacterial dermatitis is common disease that is necessary to treat with antibiotics. In recent, antibiotic-resistant bacteria is being increased in worldwide. The purpose of the present study was to evaluate the prevalence of resistant genes in Staphylococcus (S.) pseudintermedius isolated from dogs, and to compare the resistant gene profile with the result of antibiotic disc diffusion test. A total of seven S. pseudintermedius was included in the study. Bacterial identification was performed by 16S ribosomal RNA gene sequence analysis. S. pseudintermedius isolates had more than one antibiotic resistant gene (mecA, blaZ and aac(6')/aph(2"). While all isolates were PCR positive to blaZ gene, only two isolates were resistant to amoxicillin/clavulanate. Among five isolates harboring gentamicin resistance, one isolate was negative to aac(6')/aph(2")-targeted PCR. Taken together, the results suggest that resistant gene-targeted PCR and disc diffusion test are complementary to detect antibiotic resistance.

Production and Characterization of Porcine Cell Lines Overexpressing Human H-Transferase (사람 H-Transferase 유전자 과발현 형질전환 체세포주 확립 및 검증)

  • Lee, Sang-Mi;Park, Hyo-Young;Kim, Hey-Min;Moon, Seung-Ju;Kang, Man-Jong
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.187-191
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    • 2007
  • This study was canted out to develop cell lines overexpressing human H-transferase (HT). One of the approaches to prevent hyperacute rejection in xenotransplantation might be the expression of human HT in porcine cells. In this study, we cloned human HT gene from HepG2 cells using RT-PCR to establish HT-overexpressing vector. The full-length cDNA of human HT was inserted into the 3' end of CMV promoter for construction of the overexpression vector pRc/CMV-hHT. Using ietPEI DNA transfection reagent, the vector was introduced into porcine ear skin fibroblasts from newborn piglets. Transfected cells were selected by treatment of $300{\mu}g/ml$ G418 for 12 days. After antibiotic selection, survived colonies with approximately 5mm in diameter were picked and analysed for transgene human HT by PCR. The colonies proven to be human HT transfectants were analysed by RT-PCR to determine their expressions or human HT. In all colonies tested, human HT mRNA was detected. This result demonstrates the establishment of porcine cell lines overexpressing human HT, and these cell lines may be used for the development of transgenic pigs for xenotransplantation.

Molecular detection of blaVIM, blaBIC, blaKPC, and blaSIM genes from isolated bacteria in retail meats (육류용 고기로부터 분자진단을 이용한 항생제내성 유전자 양상)

  • Hwang, You Jin
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.22 no.6
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    • pp.413-419
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    • 2021
  • The purpose of this study was to investigate the ability to treat and prevent infection by multiple Gram-negative bacterial pathogens as a last choice option in the treatment of serious infections in clinical settings. The global spread of extended-spectrum 𝛽-lactamases (ESBLs) and/or carbapenemases in microorganisms are of enormous concern to health services because they are often associated with multi-drug resistance which significantly restricts the antibiotic treatment options. In this study, the antimicrobial resistance profiles of bacteria isolated from South Korean market-derived meat samples were determined by the disc diffusion method. PCR was used to detect the presence of antibiotic resistance genes and ESBL producing genes. In total, we tested 181 isolated colonies from 36 market-derived meat samples. Single PCR and DNA sequencing results revealed that genes blaVIM, blaBIC, blaKPC, and blaSIM were present in the bacteria isolated from retail meat. The bacteria in the meat were separately sequenced and based on alignment, four different bacteria were identified. These findings suggest that bacteria found in retail meats are a reservoir for the spreading of ESBL blaVIM, blaBIC, blaKPC, and blaSIM resistance genes and bacteria strains.

Molecular Monitoring of Plankton Diversity in the Seonakdong River and Along the Coast of Namhae (분자 모니터링을 이용한 서낙동강과 남해 연안 플랑크톤 군집 분석)

  • Kim, Bo-Kyung;Lee, Sang-Rae;Lee, Jin-Ae;Chung, Ik-Kyo
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.15 no.1
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    • pp.25-35
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    • 2010
  • The biodiversity of eukaryotic plankton has commonly been used to evaluate the status of aquatic ecosystems. Therefore, an accurate and rapid method for species identification is needed to reveal the biodiversity of environmental water samples. To date, molecular methods have provided a great deal of information that has enabled identification of the hidden biodiversity in environmental samples. In this study, we utilized environmental polymerase chain reaction (PCR) and constructed the 18S nuclear ribosomal RNA clone library from environmental water samples in order to develop more efficient methods for species identification. For the molecular analysis, water samples were collected from the Seonakdong River (Gimhae Bridge) and the coast of Namhae,(Namhaedo). Colony PCR and restriction fragment length polymorphism of PCR (PCR-RFLP) were then adopted to isolate unique clones from the 18S rDNA clone library. Restriction fragment length polymorphism pattern analysis of the Gimhae Bridge sample revealed 44 unique clones from a total of 60 randomly selected clones, while analysis of the Namhae sample revealed 27 unique clones from 150 clones selected at random. A BLAST search and subsequent phylogenetic analysis conducted using the sequences of these clones revealed hidden biodiversity containing a wide range of taxonomic groups (Heterokontophyta (7), Ciliophora (23), Dinophyta (1), Chytridiomycota (1), Rotifera (1) and Arthropoda (11) in the Gimhae Bridge samples Ciliophora (4), Dinophyta (3), Cryptophyta (1), Arthropoda (19) in the Namhae samples). Therefore, the molecular monitoring method developed here can provide additional information regarding the biodiversity and community structure of eukaryotic plankton in environmental samples and helps construct a useful database of biodiversity for aquatic ecosystems.

Isolation and Phylogenetic Analysis of Orchid Rotting Fungus, Fusarium sp. KS-01 (난 썩음병균 Fusarium sp. KS-01의 분리 및 계통학적 분석)

  • Park, In-Jae;Shin, Kwang-Soo
    • The Korean Journal of Mycology
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    • v.33 no.2
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    • pp.92-94
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    • 2005
  • A orchid rotting fungus was isolated and identified. The isolate was consistent with the genus Fusarium in morphological and cultural characteristics. The partial 18S rRNA sequence of the isolate showed high similarity with anamorph or telemorph of Fusarium and other Fusarium species. In phylogenetic analysis, the isolates was poorly related to other Fusarium species. The isolate closely related to Fusarium sp. LP-A2/3.

유류오염 토양의 화학.생물학적 통합처리 과정 중의 미생물 군집 변화

  • Choi Jeong-Hye;Bae Jae-Sang;Park Yeon-Jeong;Kim Su-Gon;Go Seong-Cheol
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2006.04a
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    • pp.29-32
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    • 2006
  • 화학적 산화처리와 bioremedation 기법을 개별적 또는 복합적으로 동시에 적용함으로써 한 개별 기법의 단점을 보완하고 현장적용성을 증대시킬 수 있는 통합기법을 개발하고자 하였다. 펜톤유사 반응을 통해 고농도의 유류를 산화분해 시킨 후 미생물 처리를 통해 잔류 유류 오염물질을 제거하고자 하였다. 유류 오염토양의 화학 생물학적 통합처리 공정의 현장 적용성 및 토양 미생물에 미치는 영향을 검증하기 위해 처리과정 전 후의 미생물 군집구조를 분석하였다. 또한 토양 내 유류 분해균을 분리하기 위해 탄소원으로 경유와 벙커C를 이용하여 농화배양을 수행하였다. 경유 분해균 10여종, 벙커 C 분해균 6종을 분리하여 분해능 및 동정을 시도하였다. 또한 유류 분해미생물의 consortia를 분자생물학적 기법으로 분석을 시도하였다.

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Occurrence of Plant Parasitic Nematodes in Chrysanthemum and ITS and D3-28S rDNA Characterization of Pratylenchus spp. (국화재배지의 식물기생선충 분포조사 및 뿌리썩이선층의 ITS와 D3-28S rDNA 특성조사)

  • Han, Hye-Rim;Lee, Jae-Kook;Choi, Dong-Ro;Han, Man-Jong;Park, Byeong-Yong
    • Korean journal of applied entomology
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    • v.45 no.3 s.144
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    • pp.293-299
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    • 2006
  • A survey was conducted to find out the major plant parasitic nematode in Chrysanthemum morifolium fields in Korea from May to June in 2005. A genus of Pratylenchus was determined as the most important plant parasitic nematode based on analysis of total 50 samples from 8 cities of chrysanthemum field. Pratylenchus showed 86% occurrence rate and average numbered 1,095 per 200cc soils and 1g root. Five Pratylenchus isolates, 'Muan', 'Masan', 'Tean', 'Gumi', 'Jeongup', were selected for the molecular identification of the species of Pratylenchus, and ITS and D3-28S ribosomal DNA were amplified by PCR. For the ITS, only 'Muan' isolate was differentiated by total 1 kb PCR amplification, which was 200 bp larger than all the other isolates. There was no size variation in amplified D3-28S rDNA and all isolate represented approximately 320 bp of PCR product. Sequence data of D3-28S rDNA were analysed by MegAlign program in DNASTAR software and phylogenetic tree was constructed. Sequence homology was 100% between 'Gumi' isolate and 'Tean' isolate and 'Jeongup' isolate was also close to these isolates by 99.7% sequence homology. 'Gumi', 'Tean' group and 'Jeongup' isolate were determined to be closely related to Pratylenchus vulnus by 96.7% and 96.3% similarity in respectively. D3 sequence of 'Masan' isolate was 100% identical to P. penetrans, and 'Muan' isolate showed 99.7% similarity to P. brachyurus. This result was congruent with the branch divergence pattern shown in phylogenetic tree.

Isolation of Yeasts from Wild Flowers in Gyonggi-do Province and Jeju Island in Korea and the Production of Anti-gout Xanthine Oxidase Inhibitor (경기도와 제주도 야생화들로부터 효모의 분리, 동정 및 항통풍성 Xanthine oxidase 저해물질의 생산)

  • Hyun, Se Hee;Mun, Hye Yeon;Lee, Hyang Burm;Kim, Ha-Kun;Lee, Jong-Soo
    • Microbiology and Biotechnology Letters
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    • v.41 no.4
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    • pp.383-390
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    • 2013
  • Several yeasts were isolated from flowers found in Gyonggi-do Province and Jeju island in Korea. They were then identified by a comparison of their PCR-amplified D1/D2 regions of 26S rDNA, internal transcribed spacer 1 and 2 inclusive of 5.8S rDNA, using the BLAST database. A total of fifty four yeast strains were isolated from wild flowers in Gyonggi-do and the genus Pseudozyma was noted as being dominant. A total of thirty two strains were isolated from Songaksan and Seongsan-ilchulbong in Jeju island and Sporobolomyces ruberrimus was seen to be dominant. The anti-gout xanthine oxidase inhibitory activities of the culture broths and cell-free extracts from eighty six yeast strains were then determined. The cell-free extracts of Pseudozyma hubeiensis 228-S-1 exhibited the highest xanthine oxidase inhibitory activity of 19.6%. The XOD inhibitor was also maximally produced when Pseudozyma hubeiensis 228-S-1 was cultured at $30^{\circ}C$ for 36h in YEPD medium.

Identification of Korean Poaceae Weeds Based on DNA Sequences (DNA 염기서열에 기초한 벼과 잡초의 분자생물학적 동정)

  • Lee, Jeongran;Kim, Chang-Seok;Lee, In-Yong;Oh, Hyun-Ju;Kim, Jung Hyun;Kim, Sun Yu
    • Weed & Turfgrass Science
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    • v.4 no.1
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    • pp.26-34
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    • 2015
  • Korean Poaceae includes approximately 80 species of the agricultural weeds. Precise species identification is the first step for more effective weed management in the agricultural fields. However, the identification of species in Poaceae is not easy without the assistance of taxonomists or identification experts although they are relatively easy to distinguish from the plants of the other family by the unique characteristics of caryopsis. Thus, DNA barcode was suggested as an alternative powerful technique for species identification by using short sections of DNA from a specific region of the genome. Two standard barcode markers of vascular plants, chloroplast rbcL and matK, and a supplementary nuclear ribosomal Internal Transcribed Spacer (ITS) region were used for barcode of major Korean Poaceae weeds, 403 individuals of 84 taxa. All the barcode markers revealed a good level of sequencing success with the lowest 73.7% for matK and the highest 88.8% for rbcL. The barcode sequences were deposited to the National Center for Biotechnology Information (NCBI) database for public use. Combined matK and ITS showed very high resolving power with 92.9%. Besides the identification of weeds for weed managment, the generated DNA barcode data could be used for many other applications such as rapid biodiversity assessment and conservation prioritization.