• 한국미생물·생명공학회지
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• 제50권2호
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• pp.301-309
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• 2022

본 연구에서는 제럼본에 처리에 의해 유도된 H. pylori 유전자의 전사적 변화를 분석하였다. NGS를 사용하여 RNA 발현 변화를 분석한 다음 그 결과를 검증하기 위해 RT-PCR을 수행하였다. NGS 분석 결과, 1,632개의 유전자 중 총 23개가 제럼본 처리에 의해 유의하게 발현이 변화된 특이발현 유전자로 분석되었다. DNA 복제와 전사, 병원성 인자 및 T4SS 성분과 관련된 유전자 중 10개는 현저하게 하향 조절되었고 5개는 상향 조절되었다. RT-PCR을 이용하여 유전자의 발현 수준을 재확인하였고 그 결과, 14개 유전자에서 NGS와 동일하게 발현 양상이 변화하였다. RT-PCR은 제럼본 처리에 의해 10개의 유전자(dnaE, dnaQ, rpoA, rpoD, secA, flgE, flhA, virB5, virB8, virB9)의 발현 감소와 4개의 유전자(flaA, flaB, virB4, virD4)의 발현 증가를 보였다. 이러한 본 연구의 결과는 제럼본이 다양한 H. pylori의 병원성과 관련된 인자들을 조절함으로써 H. pylori 감염의 잠재적인 치료제가 될 수 있음을 시사한다.

• 대한전기학회:학술대회논문집
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• 대한전기학회 1994년도 추계학술대회 논문집 학회본부
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• pp.342-344
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• 1994

This paper presents a design technique of SOFLC(Self -Organizing Fuzzy Logic Controller). It is composed of three parts: FLC(Fuzzy Logic Controller) part, RPO (Repeat Parameter Organizing) part, and RTPO (Real Time Parameter Organizing) part. The FLC part is controlled by initial parameters ($a_1$, $a_2$, $a_3$, $b_1$, $b_2$, $b_3$) the RPO part improves parameters by evaluating the performance of control responses controlled by FLC, and the RTPO organizes the parameters for real time in order to have the same value of the control response($y_k$) and the target response($y_k\;^*$).

• International Journal of Oral Biology
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• 제36권1호
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• pp.1-6
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• 2011

The purpose of this study was to develop species-specific real-time quantitative PCR (RT-qPCR) primers for use in the detection of Aggregatibacter actinomycetemcomitans. These primers were designed based on the nucleotide sequences of the RNA polymerase ${\beta}$-subunit gene (rpoB). We assessed the specificity of the primers against nine strains of A. actinomycetemcomitans, eight strains (three species) of the Haemophilus genus, and 40 strains of 40 other oral bacterial species. Primer sensitivity was determined by testing serial dilutions of the purified genomic DNAs of A. actinomycetemcomitans ATCC $33384^T$. Our data reveal that we had obtained species-specific amplicons for all of the tested A. actinomycetemcomitans strains, and that none of these amplicons occurred in any of the other species. Our PCR protocol proved able to detect as little as 2 fg of A. actinomycetemcomitans chromosomal DNA. Our findings suggest that these qRT-PCR primers are suitable for application in epidemiological studies.

• International Journal of Oral Biology
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• 제41권3호
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• pp.149-154
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• 2016

The purpose of this study was to develop Streptococcus sobrinus-specific qPCR primers based on the nucleotide sequence of the RNA polymerase ${\beta}-subunit$ gene (rpoB). The specificity of the primers was determined by conventional polymerase chain reaction (PCR) with 12 strains of S. sobrinus and 50 strains (50 species) of non-S. sobrinus bacteria. The sensitivity of the primers was determined by quantitative real-time PCR (qPCR) with serial dilutions of the purified genomic DNAs (40 ng to 4 fg) of S. sobrinus ATCC $33478^T$. The specificity data showed that the S. sobrinus-specific qPCR primers (RTSsob-F4/RTSsob-R4) detected only the genomic DNAs of S. sobrinus strains with a detection limit of up to 4 fg of S. sobrinus genomic DNA. Our results suggest that the RTSsob-F4/RTSsob-R4 primers are useful in detecting S. sobrinus with high sensitivity and specificity for epidemiological studies of dental caries..

• 대한임상검사과학회지
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• 제41권1호
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• pp.24-30
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• 2009

Rifampicin (RIF) and isoniazid (INH) are the most important drug for the treatment of Mycobacterium tuberculosis. Mutations correlated to rifampicin and isoniazid-resistance have been detected in rpoB gene and katG gene, respectively. Of the rifampicin-resistant isolates, 90% showed mutations in rpoB gene at codon 507 to 533. Isoniazid-resistant isolates analysed had a mutation in katG at codon 315. The aim of this study is to develop a pyrosequencing-based approach for rapid detection of ripampin or isoniazid resistant M. tuberculosis based on characterization of all possible mutation in the target region. For this study, the DNA selected from 35 cases of MTB PCR positive clinical sample such as bronchial washing, sputum, and pleural fluid. RIF or INH resistant was analyzed by pyrosequencing data of rpoB and katG gene. 28 (80%) and 7 (20%) of 35 MTB PCR positive DNAs were occured rifampicin-sensitivity and resistant, respectively. For INH, 30 (85.7%) and 5 (14.5%) cases were detected isoniazid-sensitivity and resistant, respectively. When pyrosequencing analysis was compared with ABI sequencing analysis, both analysis were presented same result, but pyrosequencing analysis was more rapid than ABI sequencing analysis. In conclusion, we found that pyrosequencing technology offers high accuracy, specificity, short turn around time and a high throughput in detection of rifampicin or isoniazid resistance in M. tuberculosis.

• International Journal of Oral Biology
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• 제35권1호
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• pp.13-19
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• 2010

The DNA probes Pn17 and Pn34 were evaluated for their ability to specifically detect clinical strains of P. intermedia and P. nigrescens from a Korean population by dot blot hybridization. These probes were sequenced by extension termination and their specificity was determined by Southern blot analysis. The results revealed that the Pn17 sequence (2,517 bp) partially encodes an RNA polymerase beta subunit (rpoB) and that Pn34 (1,918 bp) partially encodes both rpoB (1-169 nts) and the RNA polymerase beta subunit (rpoB'; 695-1918 nts). These probes hybridized with both HindIII- and PstI-digested genomic DNAs from the strains of P. intermedia and P. nigrescens used in this study. Interestingly, each of the hybrid bands generated from the HindIII-digested genomic DNAs of the two bacterial species could be used to distinguish between them via restriction fragment length polymorphism. These results thus indicate that Pn17 and Pn34 can simultaneously detect P. intermedia and P. nigrescens.

• Annals of Clinical Microbiology
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• 제17권1호
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• pp.23-27
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• 2014

신속발육항산균은 자연환경에서 흔히 검출되는 균으로, 기회감염병원체로 인체감염의 주요 원인균으로서 인식되고 있다. M. conceptionense는 분자역학적 분석을 통해 M. fortuitum의 세번째 생체변이종에서 분리되어 새로운 종으로 명명되었다. 그러나 이 균에 의한 감염 보고는 많지 않고 특히 수술후 창상 감염에 대한 보고는 거의 없다. 이에 저자들은 16S rRNA, hsp65 및 rpoB 유전자 염기순서분석을 이용하여 동정한 M. conceptionense에 의한 수술 후 창상감염을 보고하고자 한다.

• Tuberculosis and Respiratory Diseases
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• 제46권5호
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• pp.618-627
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• 1999

연구배경 : Rifampicin (RFP)은 결핵단기요법의 근간이 되는 중요한 약제이다. RFP내성 결핵균은 대부분 isoniazid (INH)에도 내성이며, 따라서 RFP내성은 다제약제내성의 지표이다. 그러나 최근 서구에서는 드문 것으로 알려진 RFP단독내성결핵이, 특히 HIV감염과 연관되어 많은 빈도로 보고되었다. 따라서 서구와는 다른 환경을 가진 국내에서 RFP단독내성결핵의 빈도, 가능한 원인들, 및 임상상을 알아보고자 하였다. 대상 및 방법 : 1990년 1월부터 1997년 8월까지 서울중앙병원에서 대한결핵협회에 의뢰하여 결핵균 감수성검사 결과가 확인된 699명(921검체)중 INH감수성, RFP내성결과를 보인 총 18명(31검체)을 대상으로 의무기록을 검토하였다. 과거결핵의 병력, 약제복용력, 동반된 전신질환 유무, HIV검사 여부, 위장질환 여부, 감수성 결과의 변화 양상, 그리고 RFP단독내성 확인후 약제의 변경등 임상 경과를 고찰하였다. 또한 보관된 6 균주는 염기서열결정법으로 rpoB유전자의 돌연변이 양상을 확인하였다. 결 과 : 18명의 평균나이는 $43{\pm}14$세였고 남녀비는 12 : 6이었다. 18명중 16명(89%)에서 과거 결핵을 앓은 병력이 있었고, 이 중 약제복용력이 확인된 12명중 11명은 불규칙하게 약제를 복용하였다. 특기할 만한 위장질환을 진단받은 병력은 없었으며, RFP을 단독으로 복용한 병력도 없었다. 기저질환으로 당뇨병이 4예에서 있었다. HIV검사가 시행된 2명 모두 음성이었다. 11명에서 감수성검사가 2회 이상 반복 시행되었는데, 6명에서는 RFP단독내성을 유지하였고 5명에서는 다제내성으로 바뀌었다. 8명(44%)은 완치되었으며, 6명(33%)은 치료실패하였고 3명(17%)은 추적검사에서 소실되었으며, 1명은 아직 치료중이다. rpoB 유전자 염기서열결정법을 시행한 6균주중 5예에서 rpoB유전자 돌연변이가 발견되었으나 새로운 형태의 돌연변이는 없었다. 결 론 : HIV감염율이 서구에 비하여 낮은 국내에서 RFP단독내성 폐결핵의 임상상은 다제내성결핵과 유사하였다. 그러나 일부 RFP단독내성결핵은 RFP을 포함한 1차약제만으로도 완치가 가능하였고, 이는 RFP 단독내성이 다제내성결핵과는 다른 부류일 가능성도 시사하였다.

• The Plant Pathology Journal
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• 제34권1호
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• pp.1-10
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• 2018

A previously unreported bacterial disease on chili pepper (Capsicum annuum L.) seedlings affecting as many as 4% of seedlings was observed in greenhouses in Chihuahua, Mexico (Delicias and Meoqui counties). Initial lesions appeared as irregular small spots on leaves and brown necrosis at margins tips were observed. Later, the spots became necrotic with a chlorotic halo. Advanced disease was associated with defoliation. A Gram negative, rod-shaped bacterium was isolated from diseased chili pepper seedlings. Three inoculation methods revealed that isolated strains produce foliage symptoms, similar to those observed in naturally infected seedlings. Pathogenic strains that caused symptoms in inoculated seedlings were re-isolated and identified to fulfill koch's postulate. Polyphasic approaches for identification including biochemical assays (API 20E and 50CH), carbon source utilization profiling (Biolog) and 16S rDNA, hsp60 and rpoB sequence analysis were done. Enterobacter cloacae was identified as the causal agent of this outbreak on chili pepper seedlings.

• The Plant Pathology Journal
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• 제33권1호
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• pp.21-33
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• 2017

Citrus blast caused by bacterium Pseudomonas syringae is a very important disease of citrus occuring in many areas of the world, but with few data about genetic structure of the pathogen involved. Considering the above fact, this study reports genetic characterization of 43 P. syringae isolates obtained from plant tissue displaying citrus blast symptoms on mandarin (Citrus reticulata) in Montenegro, using multilocus sequence analysis of gyrB, rpoD, and gap1 gene sequences. Gene sequences from a collection of 54 reference pathotype strains of P. syringae from the Plant Associated and Environmental Microbes Database (PAMDB) was used to establish a genetic relationship with our isolates obtained from mandarin. Phylogenetic analyses of gyrB, rpoD, and gap1 gene sequences showed that P. syringae pv. syringae causes citrus blast in mandarin in Montenegro, and belongs to genomospecies 1. Genetic homogeneity of isolates suggested that the Montenegrian population might be clonal which indicates a possible common source of infection. These findings may assist in further epidemiological studies of this pathogen and for determining mandarin breeding strategies for P. syringae control.