• Title/Summary/Keyword: rpoB2

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Rapid Drug Susceptibility Testing for Isoniazid and Rifampicin by Reverse Hybridization Assay (역교잡반응법을 이용한 아이소니아지드 및 리팜피신 신속감수성검사)

  • Park, Young Kil;Yu, Hee Kyoung;Ryu, Sung Weon;Bai, Gill Han
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.5
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    • pp.440-448
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    • 2003
  • Background : Development of rapid drug susceptibility testing provides the opportunity for rapid identification of individuals with drug resistant tubercle bacilli, allowing selection of appropriate therapeutic regimens. Methods : A total of 502 drug resistant isolates were subjected to reverse blot hybridization assay to detect mutations within genes (rpoB, katG, inhA, and ahpC) associated with rifampicin (RMP) and isoniazid (INH) resistance. Results : Among the 264 RMP resistant strains ($RMP^R$) tested, the most prevalent mutation was the Ser531Leu seen in 121 strains (46%). The second common mutation occurred in 84 strains (32%) at codon 526. And 27 strains (10%) showed the mutation at codon 516. Among all 469 INH resistant strains ($INH^R$), the katG mutation was responsible for INH. The inhA mutation was present in 88 strains (19%). In 11 isolates (2%), coexisting of the katG and inhA mutations were identified. Reverse hybridization assay successfully detected over 80% of $INH^R$ and over 92% of $RMP^R$ among Korean isolates. CONCLUSION: Reverse hybridization was useful for rapid detection of $INH^R$ and $RMP^R$.

Detection Method for Identification of Pueraria mirifica (Thai kudzu) in Processed Foods (가공식품 중 태국칡(Pueraria mirifica) 혼입 판별법 개발)

  • Park, Yong-Chjun;Jin, Sang-Wook;Kim, Mi-Ra;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Lee, Sang-Jae;Han, Sang-Bae
    • Journal of Food Hygiene and Safety
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    • v.27 no.4
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    • pp.466-472
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    • 2012
  • In this study, ribulose bisphosphate carboxylase (rbcL), RNApolymeraseC (rpoC1), intergenic spacer (psbA-trnH), and second internal transcribed spacer (ITS2) as identification markers for discrimination of P. mirifica in foods were selected. To be primer design, we obtained 719 bp, 520 bp, 348 bp, and 507 bp amplicon using universal primers from selected regions of P. mirifica. The regions of rbcL, rpoC1, and psbA-trnH were not proper for design primers because of high homology about P. mirifica, P. lobata, and B. superba. But, we had designed 4 pairs of oligonucleotide primers from ITS2 gene. Predicted amplicon from P. mirifica were obtained 137 bp and 216 bp using finally designed primers SFI12-miri-6F/SFI12-miri-7R and SFI12-miri-6F/SFI12-miri-8R, respectively. The species-specific primers distinguished P. mirifica from related species were able to apply food materials and processed foods. The developed PCR method would be applicable to food safety management for illegally distributed products in markets and internet shopping malls.

Clinical Meaning of INNO-LiPA Test in the Diagnosis of Rifampin Resistant Tuberculosis (Rifampin 내성 결핵의 진단에서 INNO-LiPA 검사법의 임상적 의미)

  • Chang, Yoon Soo;Kim, Young;Lee, Chang Youl;Choi, Jong Rak;Kim, Hyung Jung;Ahn, Chul Min;Kim, Sung Kyu
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.4
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    • pp.344-352
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    • 2003
  • Background : The prevalence of multidrug resistant tuberculosis (MDR-TB), resistant to isoniazid (INH) and rifampin (RFP), was 5.3% worldwide in 1995 and its increment has raised important public health problems. Resistance to RFP, one of the key drugs in the treatment of tuberculosis, results in grim clinical outcome. Recently rapid detection of RFP-resistant mutations in rpoB gene based on PCR method has become available. This study evaluated the prevalence of RFP resistance in first diagnosed, treatment failure, and recurred patients using INNO-LiPA test, and compared the results of INNO-LiPA with those of conventional mycobacterial drug susceptibility test. Methods : Forty-six patients, who were diagnosed of pulmonary tuberculosis and had revealed positive sputum AFB smear, were enrolled in this study from 1998 to 2002. The cases were classified as one three groups; first diagnosed, treatment failure, or recurred. RFP resistance was studied using an INNO-LiPA Rif. TB kit and compared with that obtained from drug susceptibility based on M. tuberculosis culture study. Results : Twenty-one out of 46 patients were enrolled under first diagnosis of pulmonary tuberculosis, 17 under treatment failure with first line drugs, and 8 under recurrence. The positive and negative predictive values of INNO-LiPA test in diagnosis in RFP resistant tuberculosis compared with conventional mycobacterial drug susceptibility test were 85.7% and 76.0%, respectively. INNO-LiPA result revealed rpoB gene mutation in 20 (80.0%) out of 25 patients who were diagnosed as treatment failure or recurrence, but in only 4 (19.0%) out of 21 patients who were first diagnosed as pulmonary tuberculosis. Conclusion : This study showed that RFP resistance could be diagnosed rapidly and accurately using INNO-LiPA test and that this test might be helpful for choosing second line anti-mycobacterial drugs. It might be of great help in clinical diagnosis and decision when used in complimentarily with drug susceptibility test based on M. tuberculosis culture.

Occurrence of Bacterial Stem Rot of Ranunculus asiaticus Caused by Pseudomonas marginalis in Korea

  • Li, Weilan;Ten, Leonid N.;Kim, Seung-Han;Lee, Seung-Yeol;Jung, Hee-Young
    • Research in Plant Disease
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    • v.24 no.2
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    • pp.138-144
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    • 2018
  • In December 2016, stem rot symptoms were observed on Persian buttercup (Ranunculus asiaticus) plants in Chilgok, Gyeongbuk, Korea. In the early stage of the disease, several black spots appeared on the stem of infected plants. As the disease progressed, the infected stem cleaved and wilted. The causal agent was isolated from a lesion and incubated on Reasoner's 2A (R2A) agar at $25^{\circ}C$. Total genomic DNA was extracted for phylogenetic analysis. Based on the 16S rRNA gene analysis, the isolated strain was found to belong to the genus Pseudomonas. To identify the isolated bacterial strain at the species level, the nucleotide sequences of the gyrase B (gyrB) and RNA polymerase D (rpoD) genes were obtained and compared with the sequences in the GenBank database. As the result, the causal agent of the stem rot disease was identified as Pseudomonas marginalis. To determine the pathogenicity of the isolated bacterial strain, it was inoculated into the stem of healthy R. asiaticus plant, the inoculated plant showed a lesion with the same characteristics as the naturally infected plant. Based on these results, this is the first report of bacterial stem rot on R. asiaticus caused by P. marginalis in Korea.

Nontuberculous Mycobacterial Lung Disease Caused by Mycobacterium terrae in a Patient with Bronchiectasis

  • Koh, Won-Jung;Choi, Go-Eun;Lee, Nam-Yong;Shin, Sung-Jae
    • Tuberculosis and Respiratory Diseases
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    • v.72 no.2
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    • pp.173-176
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    • 2012
  • We report a rare case of lung disease caused by Mycobacterium terrae in a previously healthy woman. A 45-year-old woman was referred to our hospital due to a chronic cough with sputum. A computed tomography scan of the chest revealed bronchiolitis in conjuction with bronchiectasis in both lungs. Nontuberculous mycobacteria were identified and isolated from the bronchoalveolar lavage fluid collected from each lung. All isolates were identified as M. terrae by various molecular methods that characterized the rpoB and hsp65 gene sequences. Antibiotic therapy using clarithromycin, rifampin, and ethambutol improved the patient's condition and successfully resulted in sputum conversion.

The Genetic Organization of the Linear Mitochondrial Plasmid mlp1 from Pleurotus ostreatus NFFA2

  • Kim, Eun-Kyoung;Youn, Hye-Sook;Koo, Yong-Bom;Roe, Jung-Hye
    • Journal of Microbiology
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    • v.35 no.4
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    • pp.264-270
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    • 1997
  • The structure of plasmid mlp1, a linear 10.2kb mitochondrial plasmid of Pleurotus ostreatus NFF A2 was determined by restriction enzyme mapping and partial sequencing. The plasmid encodes at least two proteins; a putative RNA polymerase showing homology to yeast mitochondrial RNA polymerase and to viral-encoded RNA polymerases, and a putative DNA polymerase showing significant homology to the family B thpe DNA polymerases. It also contains terminal inverted repeat sequences at both ends which are longer than 274 bp. A 1.6 kb EcoRI restriction fragment of m1p1 containing the putative RNA polymerase gene did not hybridize to the nuclear or motochondrial genomes from P. ostreatus, suggesting that it may encode plasmidspecific RNA polymerase. The gene fragment also did not hybridize with the RNA polymerase gene (RPO41) from Saccaromyces cerevisiae. The relationship between genes in m1p1 and those in another linear plasmid pC1K1 of Claviceps purpurea was examined by DNA hybridization. The result indicates that the genes for DNA and RNA polymerases are not closely related with those in C. purpurea.

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Analysis of Leuconostoc citreum strains using multilocus sequence typing

  • Sharma, Anshul;Kaur, Jasmine;Lee, Sulhee;Park, Young-Seo
    • Food Science and Biotechnology
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    • v.27 no.6
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    • pp.1755-1760
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    • 2018
  • The objective of this study was to perform genetic diversity analysis of 13 strains isolated from South Korean foods by multilocus sequence typing (MLST). For typing, seven housekeeping loci (atpA, dnaA, dnaK, gyrB, pheS, pyrG, and rpoA) were selected, amplified and analyzed. Fifty-one polymorphic sites varying from 1 to 22 in each species were identified. Thirteen sequence types were generated with allele numbers ranged from 2 to 10. The overall relationship between strains was assessed by unweighted pair group method with arithmetic mean dendrogram and minimum spanning tree. In addition, combined spits tree analysis revealed intragenic recombination. No clear relationship was observed between the isolation sources and strains. The developed MLST scheme enhanced our knowledge of the population diversity of Leu. citreum strains and will be used further for the selection of industrially important strain.

Effect of Carbon Couch Side Rail and Vac-lok In case of Lung RPO irradiation (Lung RPO 선량전달시, Carbon Couch Side Rail과 Vac-lok이 미치는 영향)

  • Kim, Seok Min;Gwak, Geun Tak;Lee, Seung Hun;Kim, Jung Soo;Kwon, Hyoung Cheol;Kim, Yang Su;Lee, Sun Young
    • The Journal of Korean Society for Radiation Therapy
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    • v.30 no.1_2
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    • pp.27-34
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    • 2018
  • Purpose : To evaluate the effect of carbon couch side rail and vacuum immobilization device in case of lung RPO irradiation. Materials and Methods : The 10, 20, 30 mm thickness of vac-lok's right side were obtained. To measure of doses, glass dosimeters were used and measured reference point is left lung center at the phantom. A, B, C, and D points are left, right, down, and up directions based on the center point. In the state of Side-Rail-Out, place the without vac-lok, with the thickness of 10, 20, and 30 mm vac-lok. After the glass dosimeters was inserted in center, A, B, C, and D points, 100 MU of 6 MV X-ray were irradiated to the referenced center point in the condition of $10{\times}10cm^2$ field size, SAD 100 cm, gantry angle 225, 300 MU/min dose rate. Five measurements were made for each point. In the state of Side-Rail-In, five measurement were made for each point under the same conditions. The average is measured on each of the five Side-Rail-Out and Side-Rail-In measurements. Results : In the presence of side rail, the dose reduction ratio was -11.8 %, -12.3 %, -4.1 %, -12.3 %, -7.3 % for each A, B, C, and D points. In the state of Side-Rail-Out, the dose reduction ratio for the using 10 mm thickness of vac-lok was -0.9 % than without vac-lok. The dose reduction ratio for the using 20 mm thickness of vac-lok was -2.0 %, for the using 30 mm thickness of the vac-lok was -3.0 % than without vac-lok. In the state of Side-Rail-In, the dose reduction ratio for the using 10 mm thickness of vac-lok was -1.0 % than without vac-lok. The dose reduction ratio for the using 20 mm vac-lok was -2.1 %, for the using 30 mm vac-lok was -3.0 % than without vac-lok. Based on the value of no vac-lok dose in the Side-Rail-In state, The dose reduction ratios for the using 10 mm, 20 mm and 30 mm thickness of vac-loks In the Side-Rail-Out that the center point were -12.7 %, -13.7 %, -14.2 % and -12.8 %, -13.8 %, -14.5 % respectively at point A. The dose reduction ratios for the same conditions to the B point were -4.9 %, -6.1 %, -7.1 % and -13.4 %, -14.4 %, -15.5 % respectively at point C. The dose reduction ratios for the same conditions to the D point were -8.4 %, -9.0 %, -10.4 % respectively. Conclusion : The attenuation was caused by presence of side rails and thickness of vac-lok. Pay attention to these attenuation factors, making it a more effective radiation therapy.

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Distribution of Bacterial Angular Leaf Spot of Strawberry and Characterization of Xanthomonas fragariae Strains from Korea (한국의 딸기세균모무늬병 발생분포 및 딸기세균모무늬병균 특성조사)

  • Yoon, Myung-Ju;Myung, Inn-Shik;Lee, Jae-Yeon;Kim, You-Shin;Lee, Yong-Hwan;Kim, Dae-Young;Lee, Young-Ki
    • Research in Plant Disease
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    • v.22 no.1
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    • pp.9-17
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    • 2016
  • Nationwide survey for angular leaf spot (ALS) of strawberry caused by Xanthomonas fragariae, a quarantine disease in Korea, was performed in November 2012. In the survey, ALS was observed in eighty eight farmers' fields of Sukok, Jinju and Okjong, Hadong in Gyeongnam Province, and one field in Namwon of Jeollabuk Province. The infected field of Namwon closed immediately after the disease diagnosed ALS. In detailed survey of Sukok and Okjong areas during February 2012 to January 2015, ALS occurrence decreased from 45% farmer's fields on December 2012 to 5% on January 2015, and from 38% on November 2013 to 5% on January 2015, respectively. Phenotypic characteristics of the Korean strains were similar to those of the type strain of X. fragariae. A multilocus sequence analysis of Korean strains of X. fragariae was conducted using four genes; dnaK, fyuA, gyrB, and rpoD. All the Korean strains had the same sequences of the four genes. The concatenated sequences of the Korean strains shared 100% with that of the type strain of X. fragariae. All strawberry cultivars tested were susceptible to the strains of X. fragariae two weeks after inoculation. The inoculated sites were necrosis and expanded, which were rated 4 based on evaluation of inoculation site.

Species Identification of Nontuberculous Mycobacteria (NTM) by PCR-Restriction Fragment Length Polymorphism (PRA) of the rpoB Gene from Three Hospitals of Busan-Kyeongnam Area

  • Choi, Sung-Ran;Kang, Min-Jung;Park, Gyu-Hwan;Kim, Da-Hye;Jeong, Da-Woon;Seo, Eun-Hye;Lee, Hyang-Min;Park, Hyun-Kyung;Jeong, Jin-Yee;Lee, Jung-Min;Jeong, Soo-Young;Lee, Jun-Young;Cho, Eun-Jin;Jekal, Suk;Kim, Chung-Hwan
    • Korean Journal of Clinical Laboratory Science
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    • v.45 no.2
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    • pp.48-53
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    • 2013
  • Recently, the isolation rate of nontuberculous mycobacteria (NTM) in clinical laboratories and the incidence of NTM infections are on the increase in Korea, but there have been only a few studies that reveal the general aspect of NTM isolation or species distribution. Therefore, this study was performed to examine the species identification by PCR-restriction fragment length polymorphism analysis (PRA, PCR-RFLP), and the clinical significance of mycobacterial cultures. PRA was used during the novel region of the rpoB gene and was developed for rapid and precise identification of mycobacteria to the species level. From January 2012 to April 2012, we examined pre-identified nontuberculous mycobacteria (60 species in 3 hospital of Busan-Kyeongnam area). We confirmed 4 (6.6%) Mycobacterium tuberculosis (MTB) and 56 (93.4%) NTM from 60 pre-identified NTM species by multiplex PCR (MolecuTech $MTB-ID^R$ V3, YD Diagnostics, Korea) and PRA (Myco-ID, YD Diagnostics, Korea). The distribution of 56 NTM species were M. intracellulare type I 15 (26.7%), M. avium 14 (25%), M. abscessus 11 (19.5%), M. kansasii type I 3 (5.4%), M. pulveris 2 (3.6%), M. intracellulare type, M. chelonae, M. kansasii type V, M. gallinarum, M. wolinskyi. Respectively, 1 (1.8%) and 6 (10.7%) species were not identified.

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