• Tuberculosis and Respiratory Diseases
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• v.69 no.5
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• pp.331-336
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• 2010

Background: Recently, the rate of infections with non-tuberculous mycobacteria (NTM) has been increasing in Korea. Precise identification of NTM is critical to determination of the pathogen and to target treatment of NTM patients. Methods: Sixty-eight unclassified mycobacteria isolates by rpoB PCR-RFLP assay (PRA) collected in 2008 were analyzed by National Center for Biotechnology Information (NCBI) Basic Local Alignment Search Tool (BLAST) search after sequencing of 16S rRNA, hsp65, rpoB genes. Results: Nineteen strains of 68 isolates were specified as species after sequencing analysis of 3 gene types. We found 3 M. lentifulavum, 5 M. arupense, 4 M. triviale, 4 M. parascrofulaceum, and one M. obuense. One M. tuberculosis and another M. peregrinum were mutated at the Msp I recognition site needed for rpoB PRA. The remaining 49 isolates did not coincide with identical species at the 3 kinds genes. Conclusion: Sequencing analysis of 16S rRNA, hsp65, rpoB was useful for identification of NTM unclassified by rpoB PRA.

• Korean Journal of Veterinary Service
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• v.46 no.1
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• pp.29-34
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• 2023

Mycobacterium tuberculosis complex (M. tuberculosis complex) is a causative agent of contagious chronic disease in a wide range of mammalian hosts, mainly cattle, goat, pigs, wildlife, and humans. The definite diagnosis of tuberculosis is made based on culture of M. tuberculosis, but it takes a long time. In the present study, we analyzed whether the detection time of M. tuberculosis could be reduced when cultured in the medium containing the culture-promoting ingredients-107 (CPI-107) using the BacT/Alert 3D system, an automatic culture system. The time to detection (TTD) tended to decrease as the added concentration of CPI-107 increase. In the case of low numbers of M. tuberculosis, it decreased by 21.0% at 1.2 mg/mL of CPI-107 and by 15.9% in the case of high numbers of M. tuberculosis. In the culture using clinically isolated M. tuberculosis strains, the shortening of the culture time by CPI was more evident. In conclusion, the detection time of M. tuberculosis was shortened in the medium added with CPI-107, and this could be used for isolation, culture and drug susceptibility test of M. tuberculosis.

• Tuberculosis and Respiratory Diseases
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• v.45 no.6
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• pp.1123-1142
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• 1998

Background : The frequency of MOTT has risen as the prevalence of tuberculosis has been declining. Our country has been also. The most of MOTT was resistant to the major anti-tuberculous drugs. Method : To compare clinical characteristics and frequencies of MDR tuberculosis with MOTT, the author studied 65 patients showing AFB culture positive with sputum. The data were collected from 176 patients who had been admitted at the National Masan Tuberculosis Hospital from May to June, 1997 to April, 1998. Result : The frequency of MDR tuberculosis was 43.1% and that of MOTT was 9.2%. Among 65 isolated mycobacteria, 3 cases were M. intracellulare. 2 cases were M. fortuitum, and 1 case was unidentified MOTT. The most frequent age group in 65 culture positive patients was 4th decade and the mean age was 44. The mean age was 61 in MOTT and 42 in M. tuberculosis and had significant difference(p<0.01). The numbers with past history of treatment were 2.3 in MDR tuberculosis and 1.7 in non-MDR tuberculosis and had significant difference(p<0.05). At the time of admission, the most frequent regimen for the treatment of MDR tuberculosis was 24 months regimen(85.7%) with the 2nd line anti-tuberculosis drugs. For non-MDR tuberculosis, 9 or 12 months regimen (72.9%) with the 1st line anti-tuberculosis drugs and had significant difference (p<0.01). At the time of admission, the symptom of weight loss was shown in 84.7% of M. tuberculosis and 50.0% in MOTT and there was significant difference(p<0.05) between them. All of the MOTT were identified to be resistant against INH and PAS. Drug resistance rates to INH, OFX(p<0.01) and PAS(p<0.05) in MOTT were higher than in MDR. All of three M. intracellulare strains were resistant to INH, RFP, PAS and OFX. All of two M. fortuitum strains were resistant to most anti-tuberculosis drugs. And the other MOTT was resistant to INH, EMB and PAS. Conclusion : MOTT was more common in elderly patients than M. tuberculosis. MOTT cases should be considered to be the probability of multiple drug resistance and treatment failure during the 1st treatment because they showed more resistance to anti-tuberculosis drugs than M. tuberculosis cases. Therefore, there should be more careful investigations for clinical characteristics, natural history of disease, and efficient management for MOTT.

• Biomedical Science Letters
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• v.10 no.2
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• pp.163-169
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• 2004

Worldwide, tuberculosis remains one of the leading infectious diseases, accounting for nearly 3 million deaths and more than 8 million new cases annually. DNA typing of Mycobacterium tuberculosis is important for the control of tuberculosis, since it can be used to track transmission route of tuberculosis, source of internal laboratory contaminations, and to answer questions on the nature of tuberculosis infections such as reactivation or exogenous reinfection of disease. At present, IS6110-based RFLP is the choice of method for typing large numbers of clinical isolates of M. tuberculosis, since it has the highest resolution power. However, RFLP requires long time, high cost and qualified experts, so only reference level laboratories can use the RFLP technique. In order to have an optional molecular typing method suitable for the clinical settings, this study evaluated the use of one of PCR-based typing methods, IS6110-based outward PCR for typing clinical isolates of M. tuberculosis. In brief, the results from this study showed that IS6110-based RFLP is useful to discriminate diverse clinical isolates of M. tuberculosis as well as to identify clinical isolates that belong to the same family or cluster groups that have been previously classified by RFLP analysis. In addition, the banding profiles resulted from IS6110-based outward PCR seemed to represent genomic characteristics of M. tuberculosis, since strains belong to the K-family generated unique band that is not present in any other strains but present only in the genome of K-family strains. The IS6110-based outward PCR was also shown to be useful with DNAs isolated directly from liquid cultures indicating this method can be suitable for typing M. tuberculosis in clinical settings.

• Korean Journal of Veterinary Service
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• v.36 no.2
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• pp.79-86
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• 2013

Mycobacterium (M.) bovis, a member of the M. tuberculosis complex (MTC), is a re-emerging, zoonotic agent of bovine tuberculosis whose prevalence probably depends on variations in direct exposure to cattle and ingestion of raw milk. Accurate species differentiation of M. bovis and M. tuberculosis is needed to distinguish between human and zoonotic tuberculosis. This study successfully developed a loop-mediated isothermal amplification (LAMP) assay for rapid detection and differentiation of M. bovis and M. tuberculosis, however showed negative reactions in eight non-tuberculous mycobacteria (NTM) samples and ten other bacterial species. Sensitivity of this assay for detection of genomic M. bovis DNA was 10 $fg/{\mu}l$. And this assay successfully detected M. bovis in bovine clinical specimens. In conclusion, the LAMP assay is a simple and powerful tool for rapid detection of M. bovis in both pure bacterial culture and in clinical samples.

• Korean Journal of Veterinary Service
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• v.34 no.4
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• pp.321-331
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• 2011

Mycobacterium bovis, a member of the M. tuberculosis complex (MTC), is the causative agent of bovine tuberculosis. Detection of M. bovis and M. tuberculosis using conventional culture- and biochemical-based assays is time-consuming. Therefore, a simple and sensitive molecular assay for rapid detection would be of great help in specific situations such as faster diagnosis of bovine tuberculosis (bTB) infection in the abattoirs. We developed a novel multiplex real-time PCR assay which was applied directly to biological samples with evidence of bTB and it was allowed to differentiate between M. bovis and M. tuberculosis. The primers and TaqMan probes were designed to target the IS1081 gene, the multi-copy insertion element in the MTC and the 12.7-kb fragment which presents in M. tuberculosis, not in the M. bovis genome. The assay was optimized and validated by testing 10 species of mycobacteria including M. bovis and M. tuberculosis, and 10 other bacterial species such as Escherichia coli, and cattle lymph nodes (n=113). The tests identified 96.4% (27/28) as M. bovis from the MTC-positive bTB samples using conventional PCR for specific insertion elements IS1081. And MTC-negative bTB samples (n=85) were tested using conventional PCR and the real-time PCR. When comparative analyses were conducted on all bovine samples, using conventional PCR as the gold standard, the relative accuracy of real-time PCR was 99.1%, the relative specificity was 100%, and the agreement quotient (kappa) was 0.976. The detection limits of the real-time PCR assays for M. bovis and M. tuberculosis genomic DNA were 10 fg and 0.1 pg per PCR reaction, respectively. Consequently, this multiplex real-time PCR assay is a useful diagnotic tool for the identification of MTC and differentiation of M. bovis and M. tuberculosis, as well as the epidemiologic surveillance of animals slaughtered in abattoir.

• The Journal of the Korean Society for Microbiology
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• v.17 no.1
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• pp.87-93
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• 1982

Two cases of pulmonary disease in a 54 year-old female and a 70 year-old male patient due to Mycobacterium avium-intracellulare complex(MAIC) and a case of pulmonary infection ina 69 year-old male patient due to M. fortuitum(MF) were found recently in this institute. All three patients had a long history of anti-tuberculous chemotherapy because they were initially diagnosed as pulmonary tuberculosis. A 70 year-old male patient infected with MAIC had an unsuccessful chemotherapy history of isoniazid(INH), para-aminosalicylic acid(PAS) and streptomycin(SM) with an incomplete, temporary, symptomatic improvement, for three years since 1964 when he was first diagnosed as pulmonary tuberculosis on physical examination. A 54 year-old female patient infected with MAIC also had an unsuccessful chemotherapy history with the various anti-tuberculous drugs since 1958. Both patients discharged large number of MAIC in their sputum specimens for at least more than one year, but no M. tuberculosis at all. A 69 year-old male patient infected with MF was diagnosed as moderately advanced pulmonary tuberculsis in 1977. Combined chemotherapy with INH+PAS+pyrazinamide(PZA) improved his clinical symptoms, however, his chest radiograph was deteriorated again in 1980 one year after he stopped therapy. Therefore he started chemotherapy again with INH+ethionamide(TH)+cycloserine(CS) but no improvement was noticed. MF was cultured from his sputum in August 1981 and he continuously discharged the same bacilli until last examination of January 1982. Whether all three patients were initially !infected with nontuberculous mycobacteria or complicated with predisposing tuberculosis was not clear because there were no reliable bacteriological examination records.

• The Journal of Internal Korean Medicine
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• v.21 no.4
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• pp.555-563
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• 2000

Objective : In order to know the antibacterial effects of Kyungok-go against Mycobacteria tuberculosis, Methods : In this study, I inverstigated these effects in terms of combination of other antibiotics with and without Kyungok-go on several different media conditions. Results: On Ogawa medium, Kyungok-go of the $10{\mu}/ml$ concentration showed the anti-Mycobacteria tuberculosis activity against antibiotic drug-sensitive strain. On Middle-blue medium, Kyungok-go of the $10{\mu}/ml$ concentration showed the anti-Mycobacteria tuberculosis activity against antibiotic drug-sensitive strain. Kyungok-go showed the anti mycobacteria tuberculosis activity with the meaningful result above a certain concentration. The resistance against M, tuberculosis as the concentration of Kyungok-go was decreased significantlly on the high concentration($500{\mu}/ml$) When rifampicin and Kyungok-go were used together, the resistance was decreased with the statistical significance as to the persistant antibacterial effect against M. tuberculosis, When ciprofloxacin and Kyungok-go were used together, the resistance was decreased with the statistical significance as to the persistant antibacterial effect against M. tuberculosis, The combination of treatment, Kyungok-go with both rifampicin and ciprofloxacin, showed much better antibacterial effect against M, tuberculosis than antibiotics alone. Conclusions : This study shows that Kyungok-go has antibacterial effect against M. tuberculosis and in the combination of treatment, Kyungok-go with antibiotics, showed much better antibacterial effect against M. tuberculosis than antibiotics alone,.

• Journal of Microbiology and Biotechnology
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• v.31 no.12
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• pp.1632-1642
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• 2021

Tuberculosis is a highly contagious disease caused by Mycobacterium tuberculosis. It affects about 10 million people each year and is still one of the leading causes of death worldwide. About 2 to 3 billion people (equivalent to 1 in 3 people in the world) are infected with latent tuberculosis. Moreover, as the number of multidrug-resistant, extensively drug-resistant, and totally drug-resistant strains of M. tuberculosis continues to increase, there is an urgent need to develop new anti-tuberculosis drugs that are different from existing drugs to combat antibiotic-resistant M. tuberculosis. Against this background, we aimed to develop new anti-tuberculosis drugs using probiotics. Here, we report the anti-tuberculosis effect of Pediococcus acidilactici PMC202 isolated from young radish kimchi, a traditional Korean fermented food. Under coculture conditions, PMC202 inhibited the growth of M. tuberculosis. In addition, PMC202 inhibited the growth of drug-sensitive and -resistant M. tuberculosis- infected macrophages at a concentration that did not show cytotoxicity and showed a synergistic effect with isoniazid. In a 2-week, repeated oral administration toxicity study using mice, PMC202 did not cause weight change or specific clinical symptoms. Furthermore, the results of 16S rRNA-based metagenomics analysis confirmed that dysbiosis was not induced in bronchoalveolar lavage fluid after oral administration of PMC202. The anti-tuberculosis effect of PMC202 was found to be related to the reduction of nitric oxide. Our findings indicate that PMC202 could be used as an anti-tuberculosis drug candidate with the potential to replace current chemical-based drugs. However, more extensive toxicity, mechanism of action, and animal efficacy studies with clinical trials are needed.

• Tuberculosis and Respiratory Diseases
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• v.39 no.5
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• pp.425-432
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• 1992

Background: The prevalence of tuberculosis in Korea decreased remarkably for the past 30 years, while for at least the recent 10 years, the frequency of disease attributable to mycobacteria other than tuberculosis (MOTT) began to increase both in actual numbers and in the proportion of the total burden of mycobacterioses. Method: Twenty nine cases, diagnosed as having pulmonary disease due to MOTT on the basis of repeated isolations of the relevant organisms from sputum specimens and chest X-ray findings from Jan. 1982 throught Dec. 1991 at the Chest Clinic of the Korean Institute of Tuberculosis, were reviewed in terms of clinical features and courses. Results: 1) Nineteen cases (66%) were infected with Mycobacterium avium-intracellulare, 5 cases (17%) with M. fortuitum, 4 cases (14%) with M. chelonei, and 1 cases (3%) with M. szulgai. 2) The ratio of male versus female patients was 1.9:1. 3) The peak incidence was in the fifth decade. 4) Eighteen cases (62%) had moderately advanced disease and 11 cases (38%) had far advanced disease on chest P-A film. 5) The common symptoms were cough (59%), sputum (52%), and dyspnea (34%). 6) All cases had a previous history of tuberculosis. 7) Most of the isolates were highly resistant to the major antituberculosis drugs and 48~52% showed a sensitivity to cycloserine, kanamycin or enviomycin. 8) Of 19 cases which were treated and followed-up for over 12 months, 3 cases attained negative conversion on cultures (2 M. fortuitum infections, 1 M. szulgai infection). Sixteen cases failed in attaining negative conversion on cultures. However, their clinical courses were chronic and indolent despite of persistant culture positivity. Conclusion: Treatment of these patients has been difficult because of the frequency of severe underlying conditions and the natural resistance of most of the nontuberculous mycobacteria to the presently available drugs.