• 한국식물병리학회지
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• 제1권1호
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• pp.61-66
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• 1985

벼 흰빛잎마름병균주 Q7472와 Q7502의 항혈청을 사용하여 우리나라에서 수집한 벼 흰빛잎마름병균의 혈청형을 한천gel내 확산법에 의해 분류한 결과, 71균주중 65균주가 혈정형 A형, 5균주가 B-I, 1균주가 B-II형에 속하였다. 혈청형A에 속하는 균주는 $0.1-1\%$의 $CaCl_2$용액에서 자기의집이 거의 일어나지 않았으나 혈청형 B-I과 B-II에 속하는 균주는 심한 자기의집이 일어나 혈청형과 자기의집 사이에는 밀접한 관계가 있었다. 흰빛잎마름병균은 혈청학적 방법에 의해 쉽게 검색될 뿐만 아니라 한천gel내 확산법에 의해 벼 세균성줄무늬병균인 X.campestris pv. olyzicola의 부생세균인 E. herbicola와도 쉽게 구별되었다. 흰빛잎마름병균 검출은 병반을 NaCl용액에 넣은 후에 착압하거나 $100^{\circ}C$에서 1시간 열처리 후 착압한 즙액을 한천gel내 확산법에 의해 항혈청과 반응시킴으로써 가능하였으며, 병반이 적을때는 PSA배지에서 배양한 후 사용하면 효과적이었다.

• Journal of Microbiology and Biotechnology
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• 제27권6호
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• pp.1128-1137
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• 2017

The aim of this study was to explore the accuracy and feasibility of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) in identifying bacteria from environmental sources, as compared with rpoA gene sequencing, and to evaluate the occurrence of bacteria of the genus Enterococcus in wild birds. In addition, a phyloproteomic analysis of certain Enterococcus species with spectral relationships was performed. The enterococci were isolated from 25 species of wild birds in central Europe (Poland). Proteomic (MALDI-TOF MS) and genomic (rpoA gene sequencing) methods were used to identify all the isolates. Using MALDI-TOF MS, all 54 (100%) isolates were identified as Enterococcus spp. Among these, 51 (94.4%) isolates were identified to the species level (log(score) ${\geq}2.0$), and three isolates (5.6%) were identified at a level of probable genus identification (log(score) 1.88-1.927). Phylogenetic analysis based on rpoA sequences confirmed that all enterococci had been correctly identified. Enterococcus faecalis was the most prevalent enterococcal species (50%) and Enterococcus faecium (33.3%) the second most frequent species, followed by Enterococcus hirae (9.3%), Enterococcus durans (3.7%), and Enterococcus casseliflavus (3.7%). The phyloproteomic analysis of the spectral profiles of the isolates showed that MALDI-TOF MS is able to differentiate among similar species of the genus Enterococcus.

• The Plant Pathology Journal
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• 제34권5호
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• pp.451-457
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• 2018

The Sweet potato chlorotic fleck virus (SPCFV), of the genus Carlavirus (family Betaflexiviridae), was first detected as one of several viruses infecting sweet potatoes (Ipomea batatas L.) in Korea. Out of 154 sweet potato samples collected in 2012 that were showing virus-like symptoms, 47 (31%) were infected with SPCFV, along with other viruses. The complete genome sequences of four SPCFV isolates were determined and analyzed using previously reported genome sequences. The complete genomes were found to contain 9,104-9,108 nucleotides, excluding the poly-A tail, containing six putative open reading frames (ORFs). Further, the SPCFV Korean isolates were divided into two groups (Group I and Group II) by phylogenetic analysis based on the complete nucleotide sequences; Group I and Group II had low nucleotide sequence identities of about 73%. For the first time, we determined the complete genome sequence for the Group II SPCFV isolates. The amino acid sequence identity in coat proteins (CP) between the two groups was over 90%, whereas the amino acid sequence identity in other proteins was less than 80%. In addition, SPCFV Korean isolates had a low amino acid sequence identity (61% CPs and 47% in the nucleotide-binding protein [NaBp] region) to that of Melon yellowing-associated virus (MYaV), a typical Carlavirus.

• The Plant Pathology Journal
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• 제33권3호
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• pp.307-317
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• 2017

Satsuma dwarf virus (SDV) or Citrus mosaic sadwavirus (CiMV) were not consistently detected in RTPCR assay with the primer sets based on gene of Japan isolates. SDV and CiMV isolates were distinctively divided into two groups based on phylogenetic analysis of PP2 gene cloned from 22 Korean isolates, and the Korean CiMV and SDV isolates shared 95.5-96.2% and 97.1-97.7% sequence identity with Japanese isolate, respectively. We developed PP2-1 primer set based on the PP2 gene sequence of Korean isolates to simultaneously and effectively detect SDV and CiMV. And CTLV-2013 and CTV-po primer sets were newly designed for detection of Citrus tatter leaf virus (CTLV) and Citrus tristeza virus (CTV), respectively. Using these primer sets, a new multiplex PCR assay was developed as a means to simultaneously detect 4 citrus viruses, CTV, CTLV, SDV, and CiMV. The degree of detection by the multiplex PCR were consistent with those of uniplex RT-PCR for detection of each of the viruses. Therefore, the new multiplex PCR provides an efficient method for detecting 4 citrus viruses, which will help diagnose many citrus plants at the same time. We verified that 35.2% and 72.1% of 775 trees in 155 orchards were infected with SDV or CiMV (SDV/CiMV) and CTV by the multiplex-PCR assay, respectively, and CTLV was not detected in any of the trees tested.

• Tuberculosis and Respiratory Diseases
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• 제81권3호
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• pp.222-227
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• 2018

Background: Rifampicin (RFP) is one of the principal first-line drugs used in combination chemotherapies against Mycobacterium tuberculosis, and its use has greatly shortened the duration of chemotherapy for the successful treatment of drug-susceptible tuberculosis. Compensatory mutations have been identified in rpoC that restore the fitness of RFP-resistant M. tuberculosis strains with mutations in rpoB. To investigate rpoC mutation patterns, we analyzed 93 clinical M. tuberculosis isolates from patients in South Korea. Methods: Drug-resistant mycobacterial isolates were cultured to determine their susceptibility to anti-tubercular agents. Mutations in rpoC were identified by sequencing and compared with the relevant wild-type DNA sequence. Results: In total, 93 M. tuberculosis clinical isolates were successfully cultured and tested for drug susceptibilities. They included 75 drug-resistant tuberculosis species, of which 66 were RFP-resistant strains. rpoC mutations were found in 24 of the 66 RFP-resistant isolates (36.4%). Fifteen different types of mutations, including single mutations (22/24, 91.7%) and multiple mutations (2/24, 8.3%), were identified, and 12 of these mutations are reported for the first time in this study. The most frequent mutation involved a substitution at codon 452 (nt 1356) resulting in amino acid change F452L. Conclusion: Fifteen different types of mutations were identified and were predominantly single-nucleotide substitutions (91.7%). Mutations were found only in dual isoniazid- and RFP-resistant isolates of M. tuberculosis. No mutations were identified in any of the drug-susceptible strains.

• 한국응용곤충학회지
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• 제17권1호
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• pp.15-22
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• 1978

1. "Kresek" 유발균주(K균주)는 품종에 따라서 발현에 차이가 있었으며 지상부 또는 지하부에 접종하여도 "Kresek"을 유발시킬 수 있었으나 대조균주(N균주)는 전연 유발되지 않았다. 2. 엽백의 도관직경은 밀양 23이 유신이나 통일보다 현저히 작았으며 이들간에 고도의 통계적인 유의차가 인정되었다. 3. 밀양 23의 즙액에서 K균주들은 N균주들보다 세균의 증식이 왕성하였으며 통일에서는 균주들간에 증식의 차이가 없었다. 4. 밀양 23의 즙액친서는 K균주의 Dead phase가 정종 9일째부터 N균주는 4일째부터 시작되었으며 통일에서는 모두 3일째부터 시작되었다. 5. 밀양 23즙액의 경우 pH 3.0-4.0 범위에서 다량의 침전물이 형성되었으나 동일의 증액에서는 극히 소량의 침전물을 관찰할 수가 있었다.

• 대한의생명과학회지
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• 제25권1호
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• pp.40-53
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• 2019

Of the Acinetobacter spp., A. baumannii (genospecies 2) is the most clinically significant in terms of hospital-acquired infections worldwide. It is difficult to perform Acinetobacter-related taxonomy using phenotypic characteristics and routine laboratory methods owing to clusters of closely related species. The ability to accurately identify Acinetobacter spp. is clinically important because antimicrobial susceptibility and clinical relevance differs significantly among the different genospecies. Based on the medical importance of pathogenic Acinetobacter spp., the distribution and characterization of Acinetobacter spp. isolates from 123 clinical samples was determined in the current study using four typically applied bacterial identification methods; partial rpoB gene sequencing, amplified rRNA gene restriction analysis (ARDRA) of the intergenic transcribed spacer (ITS) region of the 16~23S rRNA, the $VITEK^{(R)}$ 2 system (an automated microbial identification system) and matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). A. baumannii isolates (74.8%, 92/123) were the most common species, A. nosocomialis (10.6%, 13/123) and A. pittii isolates (7.5%, 9/123) were second and third most common strains of the A. calcoaceticus-A. baumannii (ACB) complex, respectively. A. soli (5.0%, 6/123) was the most common species of the non-ACB complex. RpoB gene sequencing and ARDRA of the ITS region were demonstrated to lead to more accurate species identification than the other methods of analysis used in this study. These results suggest that the use of rpoB genotyping and ARDRA of the ITS region is useful for the species-level identification of Acinetobacter isolates.

• 대한의생명과학회지
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• 제24권4호
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• pp.430-434
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• 2018

Candida glabrata is the second most prevalent causative agent for candidiasis following C. albicans. The opportunistic yeast, C. glabrata, is able to cause the critical bloodstream infections in hospitalized patients. Conventional identification methods for yeasts are often time consuming and labor intensive. Therefore, recent studies on sequence-based identification have been conducted. Recently, sequencing the D1/D2 domain of the large subunit ribosomal RNA gene and the internal transcribed spacers (ITS) 1 and ITS2 regions of the ribosomal DNA has proven useful for DNA-based identification of most species of fungi. In the present study, therefore, fungal ITS and D1/D2 domain regions were targeted and analyzed by DNA sequencing for the accurate identification of C. glabrata clinical isolates. A total of 102 C. glabrata clinical isolates from various clinical samples including bloodstream, catheterized urine, bile and other body fluids were used in the study. The results of the DNA sequence analysis showed that the mean standard deviation of species identity percent score between ITS and D1/D2 domain regions was $97.8%{\pm}2.9$ and $99.7%{\pm}0.46$, respectively. These results revealed that the D1/D2 domain region might be a better target for identifying C. glabrata clinical isolates based on DNA sequences than the ITS1 and ITS2 regions. However, in order to evaluate the usefulness of D1/D2 domain region for species identification of all Candida species, other Candida species such as C. albicans, C. tropicalis, C. dubliniensis, and C. krusei should be verified in further studies additionally.

• 한국식품위생안전성학회지
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• 제30권1호
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• pp.103-108
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• 2015

본 연구에서는 김치로부터 W. cibaria 0D17와 W. confusa 0D23를 생화학적 특성 분석과 16s rRNA 염기서열 분석을 통해 분리, 동정하였으며, W. cibaria 0D17와 W. confusa 0D23 배양 상등액이 L. monocytogenes에 대한 항균 효과가 있는 것으로 나타났다. 또한, 실시간 정량 PCR을 통해 W. cibaria 0D17 및 W. confusa 0D23가 L. monocytogenes를 공동 배양했을 때의 생육억제 효과를 분석한 결과, $37^{\circ}C$에서는 생육 억제 효과가 있는 것으로 확인되었다. 따라서, 김치 유래 Weissella spp.가 갖는 L. monocytogenes에 대한 항균 활성에 대한 기초 자료로 활용가능 할 것으로 판단되며, W. cibaria 0D17 및 W. confusa 0D23가 생산하는 bacteriocin 등의 항균 물질 특성에 대한 연구가 추가적으로 진행될 필요가 있을 것이다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권20호
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• pp.8883-8886
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• 2014

Background: Tetracycline is an antibiotic widely used for the treatment of Helicobacter pylori infection, but its effectiveness is decreasing due to increasing bacterial resistance. The aim of this study was to investigate the occurrence of 16S rRNA mutations associated with resistance or reduced susceptibility to tetracycline ofHelicobacter pylori by real-time PCR (RT-PCR) assays from culture. Materials and Methods: Tetracycline susceptibility and minimal inhibition concentration (MIC) was determined by the Epsilometer test (Etest) method. A LightCycler assay developed to detect these mutations was applied to DNA extracted from culture. The 16S rRNA of these isolates was sequenced and resistance-associated mutations were identified. From 104 isolates of H. pylori examined, 11 showed resistance to tetracycline. Results: LightCycler assay was applied to DNA extracted from 11 tetracycline-susceptible and 11 tetracycline resistance H. pylori isolates. In our study the sequencing of the H. pylori wild types in 16 s rRNA gene were AGA 926-928 with MIC (0.016 to $0.5{\mu}g/ml$), while the sequencing and MIC for resistant were GGA and AGC, (0.75 to $1.5{\mu}g/ml$), respectively. Also we found a novel mutation in 2 strains with $84^{\circ}C$ as their melting temperatures and exhibition of an A939C mutation. Conclusions: We conclude that real-time PCR is an excellent method for determination of H. pylori tetracycline resistance related mutations that could be used directly on biopsy specimens.