• 대한의생명과학회지
• /
• 제16권3호
• /
• pp.133-138
• /
• 2010

Despite of the increasing importance of environmental mycobacteria, detection and identification of mycobacteria from environmental sources including water have been fraught with technical difficulties. Although, several protocols to optiruize isolation of mycobacteria from water sources have been reported, standard method has not yet been established. In this study, usefulness of cetylpyridinium chloride (CPC), a cationic quaternary ammonium compound, for the isolation of environmental mycobacteria from diverse water samples was evaluated. For this, water samples from diverse water sources such as effluent water, lake water, and underground water were collected, treated with diverse concentrations of CPC, and plated on the solid agar plates. Subsequently individual colonies grown on the plates were sequence analyzed for identification of each colony. In brief, the results from this study showed that the growth of mycobacteria was enhanced by use of CPC as a pre-treatment reagent to water samples by inhibiting growth of other non-mycobacteria in water. In fact, the effect of CPC to decontaminate non-mycobacteria for isolation of mycobacteria was better than 1~4% of NaOH, which is a routinely used decontaminating reagent widely employed for culturing mycobactera from sputum specimens. Therefore, the results from this study seems to support that the CPC pre-treatment may be useful for isolation of mycobacteria from diverse sources including clinical specimens which are often contaminated with other bacteria.

• KSBB Journal
• /
• 제7권3호
• /
• pp.161-165
• /
• 1992

Microemulsions에 교차결합(crosslinking)고정화한 Microemulsions sp(NRRL B-3683)를 이용하여 소수성기질인 sitosterol로부터 AD, ADD를 합성하였다. Mycobacteria bioconversion활성도는 계면활성제의 종류와 물의 양에 의존한다. Mycobacteria는 양이온 microemulsions(CTAC-buthanol-cyclohexa-ne-water)에서 가장 높은 활성도를 갖으며, 이것은 buffer에서보다 5배 이상 효율을 갖음을 나타낸다. 음이온 microemulsions에서는 활성도가 매우 낮다. 양이온 microemulsions에서활성도는 물의 양에 따라서 변하며 물의 양이 증가함에 따라서 증가한다. 20%의 물을 포함한 microemulsions에서는 5%의 물을 포함한 microemulsions에서보다 약 배의 효율을 갖는다. 이와같이 microemulsions은 미생물을 이용한 bioconversion반응의 매질로써 효과적일 수 있다.

• Journal of Microbiology and Biotechnology
• /
• 제18권7호
• /
• pp.1207-1215
• /
• 2008

To investigate the occurrence and species diversity of mycobacteria in waters, surface water samples were collected monthly from the Han River and tap water samples at the terminal sites of the distribution system. Mycobacteria in each water sample were isolated by decontamination using cetylpyridinium chloride (CPC) and cultivation on Middlebrook 7H10 agar, and then identified by polymerase chain reaction-restriction fragment length polymorphism analysis (PRA) and sequencing of the 65-kDa heat-shock protein gene (hsp65 gene). Mycobacteria were detected in 59% of the surface water samples and 26% of the tap water samples. Over half of the 158 isolates could not be identified by hsp65 PRA and gene sequencing, and several identification discrepancies were observed between the two methods. The most frequently isolated species was Mycobacterium gordonae in surface water and M. lentiflavum in tap water. M. avium complex (MAC), the most important pathogen among environmental mycobacteria, was detected in the surface water samples but not found in the tap water samples. The result demonstrated that water is an important environmental source of mycobacteria and the combined application of hsp65 PRA and sequencing was more reliable than hsp65 PRA alone to accurately identify mycobacteria present in water.

• 대한임상검사과학회지
• /
• 제47권2호
• /
• pp.83-89
• /
• 2015

결핵(Mycobacterium tuberculosis, MTB) 감염은 아직까지 전 세계에서 높은 유병률과 사망의 주요 원인이 되고 있으며 비정형 결핵(nontuberculous mycobacteria, NTM)은 최근 후천성 면역결핍증(AIDS)이나, 종양, 이식 등으로 면역력이 저하된 환자들의 임상 검체에서 분리빈도가 증가함에 따라 분리 균주의 임상적 의의가 중요시 되고 있다. 이에 항산균 염색을 이용한 객담 도말검사는 결핵균과 비정형 결핵균을 구별할 수 없다는 제한점이 있고, 균 분리배양검사는 시간이 오래 걸린다는 단점이 있어, 본 연구에서는 이러한 제한점을 가진 기존의 검사법을 대신하여 분자생물학적 방법인 중합효소 연쇄반응(polymerase chain reaction, PCR)을 이용하여 균 분리배양 검사와 비교하여 보았다. Mycobacteria를 동정하는데 항산균 염색과 3% ogawa 배지를 이용하였고, 균 분리배양 후 M. tuberculosis를 확인하기 위해서 niacin test를 실시한 결과 집락의 DNA를 추출하여 PCR후 동정된 M. tuberculosis와 niacin 양성이 일치함을 확인할 수 있었다. 또, 이 실험에서 항산성균 염색이 2+ 이상인 객담검체와 집락검체 각각에서 DNA를 추출하여 결핵균을 동정하는 방법으로 M. tuberculosis complex에 특징적으로 존재하는 insertion sequence (IS) 6110의 특정부위인 547 bp와 285 bp 부분을 증폭한 two-tube nested polymerase chain reaction을 시행하였고, 비정형 결핵균 동정법으로는 mycobacteria에 공통적으로 존재하는 rpoB 유전자 중 일부인 360 bp 부분을 증폭한 후, 제한효소 Msp I을 첨가 PCR-restriction fragment length polymorphism (RFLP)을 시행하였으며, 결핵균과 비정형 결핵균 모두 동정율에 거의 차이가 없었다. 1+인 경우는 객담검체에서 PCR한 결과가 31.2%에서 집락검체의 PCR한 결과 93.7%까지 결핵균과 비정형결핵균의 동정율이 높아졌고, trace인 경우 객담검체에서 PCR 결과가 2%에서 집락검체에서 PCR한 결과가 97.9%까지 결핵균과 비정형 결핵균의 동정율을 높일 수 있었다. 이 실험에서 항산성균 염색 1+이하 일때 객담검체와 집락검체로 PCR을 실시하면 결핵균과 비정형 결핵균의 동정율에 차이가 있고, 배양만으로는 결핵균의 동정은 가능하지만 비정형 결핵균의 동정이 가능하지 않으므로 배양검사와 PCR 검사 모두를 병행하므로써 보다 신속하고 정확한 검사결과를 내는데 도움 될 것이다.

• IMMUNE NETWORK
• /
• 제11권5호
• /
• pp.245-252
• /
• 2011

Antimicrobial peptides/proteins are ancient and naturally-occurring antibiotics in innate immune responses in a variety of organisms. Additionally, these peptides have been recognized as important signaling molecules in regulation of both innate and adaptive immunity. During mycobacterial infection, antimicrobial peptides including cathelicidin, defensin, and hepcidin have antimicrobial activities against mycobacteria, making them promising candidates for future drug development. Additionally, antimicrobial peptides act as immunomodulators in infectious and inflammatory conditions. Multiple crucial functions of cathelicidins in antimycobacterial immune defense have been characterized not only in terms of direct killing of mycobacteria but also as innate immune regulators, i.e., in secretion of cytokines and chemokines, and mediating autophagy activation. Defensin families are also important during mycobacterial infection and contribute to antimycobacterial defense and inhibition of mycobacterial growth both in vitro and in vivo. Hepcidin, although its role in mycobacterial infection has not yet been characterized, exerts antimycobacterial effects in activated macrophages. The present review focuses on recent efforts to elucidate the roles of host defense peptides in innate immunity to mycobacteria.

• 대한미생물학회지
• /
• 제8권1호
• /
• pp.33-36
• /
• 1973

This study was carried out for the classification of mycobacteria using the thin-layer chromatography of mycobacterial lipids. Results as follows: Of the 12 strains of mycobacteria, the two spots on chromatogarphy were three strains of mycobacteria(BCG, M. tuberculosis($H_{37}J$) and M. ulcerans) and three spots on chromatography were two strains of mycobacteria(M. kansasii and M. balnei). The method of thin-layer chromatography of mycobacterial lipids was considered to capable for the use of classification of mycobacteria.

• 한국정보통신학회:학술대회논문집
• /
• 한국정보통신학회 2012년도 춘계학술대회
• /
• pp.913-915
• /
• 2012

Rifampin 내성 마이코박테리아의 RNA polymerase beta subunit gene (rpoB)의 변이를 of Rifampin-resistant Mycobacteria was analyzed using nucleotide sequence of containing rifampin 내성유전자부위인 $rif^{rr}$을 포함한 rpoB DNA (351 bp)의 염기서열 분석을 이용하여 조사하였다. 본 연구를 위해 마산국립병원과 국립결핵원으로부터 전통적 배양방법으로 rifampin 내성 마이코박테리아를 수집하여 그것들의 rpoB 유전자를 염기서열 분석을 수행하였고 최근까지 보고된 것과는 다른 변이들을 확인할 수 있었다.

• 대한미생물학회지
• /
• 제9권1호
• /
• pp.33-40
• /
• 1974

Mycobacteria의 Crocin 첨가 결핵균 배지상의 보라색 색소형성과 광선조사, 호혐기상태, Crocin의 농도 및 배양시간과의 관계를 조사하고 비정형 Mycobactia의 색소형성 특성에 따른 군분류 동정에 대하여 기술하였다. 광색정군 M. kansasii는 양성으로 음성인 M. marinum과 감별이 되었으며, 색정군 M. scrofulaceum은 음성으로서 양성인 M. aquae와, 조기발육군 M. fortuitum은 3일 배양검사에서 양성으로 M. smegmatis로 부터 본류 동정이 각각 가능하였다.

• 한국정보통신학회:학술대회논문집
• /
• 한국정보통신학회 2012년도 추계학술대회
• /
• pp.991-993
• /
• 2012

rifampin 내성 유전자 부위인 $rif^{rr}$를 포함하는 RNA polymerase beta subunit gene (rpoB) (351 bp)의 DNA 서열을 분석을 이용하여 rifampin 내성 마이코박테리아의 rpoB DNA 변이를 조사하였다. 본 연구를 위해 국립마산병원과 결핵원으로 부터 기존의 재래 배양방법으로 동정한 rifampin에 대한 내성 마이코박테리아를 수집하였다. 본 연구실에서는 수집된 rifampin 내성 마이코박테리아에서 rpoB 유전자의 DNA 서열 분석을 수행하였다. 본 연구의 분석 결과로 rifampin 내성 마이코박테리아에서 $rif^{rr}$를 포함한 rpoB의 보고되지 않은 다양한 변이들이 조사되었다.

• 대한미생물학회지
• /
• 제11권1호
• /
• pp.69-78
• /
• 1976

There have been increasing reports of mycobacterioses in man and animals caused by "atypical" or "opportunist" mycobacteria. At the presnt, "opportunist mycobacterioses" are not generally responsive to antituberculosis drugs, and therefore, create considerable problems with regard to chemotherapy and control measures. In recent years studies have been made to isolate opportunist mycobacteria from soil, house-dusts and tap-water. It seemed quite interesting to define the extent of circumstantial presence of "opportunist" mycobacteria and nocardia in the soils of school-ground of primary schools and middle-high schools. This communication is the results of pilot study to isolate and identify "opportunist" mycobacteria and nocardia from 504 soil specimens of 72 schools in Seoul City. 1. Of a total of 59 isolates from 504 soil specimens tested, 32 strains were identified as opportunist mycobacteria and 27 strains as nocardia. 2. Isolation rates of opportunist mycobacteria by the areas(of specimen collection) were as follows: 36.4% in the southern area of Han-River, 33.3% in the central area, 22.7% in the outskirt area and 16.6% in the intermediate area. There observed no apparent difference in the isolation rates both-between the areas and between primary schools and middle-high schools. However, a significant difference was noted in the isolation rates between the places of soil sampling in a given school(P<0.05), i.e., the highest was the soil of refuge heaps(15.2%), and tap-water pole area(11.1%), the school-lavatory entrance(9.7%), the school gate entrance(5.5%), and iron-bar play ground(2.7%). The soil specimens from the center of school ground and from school building entrance yielded none of mycobacterial isolates. 3. Isolation rates of nocardia by the areas were as follows: 33.3% in the central area, 31.8% in the outskirt area, 27.3% in the southern ares of Han-River and 11.1% in the intermediate area. As in the case of mycobacteral isolates, there observed no apparent differences in the isolation rates both between the areas and between primary schools and middle-high schools, but a significant difference was noted between the places of soil sampling(P<0.05), i.e., the highest was the soil of school building entrance(15.2%), and of school gate entrance(6.9%), refuge heaps(5.5,%), iron-bar play ground(4.1%), the school-lavatory entrance(2.7%) and tap-water pole area(2.7%), respectively. The soil specimens from the center of, school ground yielded none of nocardia isolates. 4. Of the 32 strains of isolated mycobacteria. 15 strains were slow-growing mycobacteria and the remaining 17 strains belonged to the rapid growers. Of the 15 slow-growers. 4 strains were M. scrofulaceum-szulgai complex, 3 M. gordonae, 4 M. terrae-triviale complex, 2 M. avium-intracellulare-xenopi complex, and 2 unclassified schotochromogens. Of the 17 strains of rapid growers, 12 were M. diernhoferi, 2 M. fortuitum-peregrinum complex, 2 M. vaccae and one M. flavescens. 5. Of the 27 strains of nocardia isolated, 11 strains were N. transvalensis, 5 N. convoluta, 5 N. erythropolis, one N. vaccinii, one N. polychromogens-paraffinae complex and 4 untypable strains of orange-pigmented nocardia spp.