• Tuberculosis and Respiratory Diseases
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• v.50 no.1
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• pp.29-41
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• 2001

Background : The appearance of multiple-drug-resistant Mycobacterium tuberculosis strains has been seriously compromising successful control of tuberculosis. Rifampin-resistance, caused by mutations in the rpoB gene, can be indicative of multiple-drug-resistance, and its detection is of great importance. The present study aimed to develop an oligonucleotide chip for accurate and convenient screening of drug-resistance. Methods : In order to detect point mutations in the rpoB gene, an oligonucleotide chip was prepared by immobilizing specific probe DNA to a microscopic slide glass by a chemical reaction. The probe DNA that was selected from the 81 bp core region of the rpoB gene was designed to have mutation sites at the center. A total of 17 mutant probes related to rifampin-resistance including 8 rifabutin-sensitive mutant probes were used in this study. For accurate determination, wild type probes were prepared for each mutation position with an equal length, which enabled a direct comparison of the hybridization intensities between the mutant and wild type. Results : Mycobacterial genomic DNA from clinical samples was tested with the oligonucleotide chip and the results were compared with those of the drug-susceptibility test in addition to sequencing and INNO-LiPA Rif. TB kit test in some cases. Out of 15 samples, the oligonucleotide chip results of 13 samples showed good agreement with the rifabutin-sensitivity results. The two samples with conflicting result also showed a discrepancy between the other tests, suggesting such possibilities as existence of mixed strains and difference in drug-sensitivity. Further verification of these samples in addition to more case studies are required before the final evaluation of the oligonucleotide chip can be made. Conlcusion : An oligonucleotide chip was developed for the detection of rpoB gene mutations related to drugsusceptibility. The results to date show the potential for using the oligonucleotide chip for accurate and convenient screening of drug-resistance to provide useful information in antituberculosis drug therapy.

• Tuberculosis and Respiratory Diseases
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• v.56 no.3
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• pp.261-268
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• 2004

Background : The resurgence of tuberculosis and outbreaks of multidrug resistant (MDR) tuberculosis have increased the emphasis for the development of new susceptibility testing of the Mycobacterium tuberculosis for the effective treatment and control of the disease. Conventional drug susceptibility testings, such as those using egg-based or agar-based media have some limits, such as the time required and difficulties in determining critical inhibitory concentrations, but these are still being used in many diagnostic laboratories because of no better lternatives, considering cost and accuracy. To overcome these limits, a rapid and simple method for new susceptibility testing, using live and dead assays, was applied for a bacterial cell viability assay to distinguish dead from live bacterial cells based on two-color fluorescence. Materials and Methods Strains : Forty strains were used in this study, 20 susceptible to all antituberculosis drugs and the other 20 resistant to the four first line antituberculosis drugs isoniazid, rifampicin, streptomycin and ethambutol. Antibiotics : The four antibiotics were dissolved in 7H9 broth to make the following solutions: $0.1{\mu}g\;isoniazid(INH)/m{\ell}$, $0.4{\mu}g\;rifampicin(RMP)/m{\ell}$, $4.0{\mu}g\;streptomycin(SM)/m{\ell}$ and $4.0{\mu}g\;ethambutol(EMB)/m{\ell}$. Results : Live and dead Mycobacterium tuberculosis cells fluoresced green and red with the acridin (Syto 9) and propidium treatments, respectively. These results are very well accorded with conventional drug susceptibility testing by proportional method on Lowensen-Jensen media (L-J) containing 4 drugs (INH, RMP, EMB and SM), showing a 93.7 % accordance rate in susceptible strains and 95% in resistant strains. Conclusion : The results of the drug susceptibility testing using the live and dead bacterial cell assay showed high accordance rates compared with the conventional proportion method on L-J. This finding suggests that the live and dead bacterial cell assay can be used as an alternative to conventional drug susceptibility testing for M. tuberculosis strains.

• Tuberculosis and Respiratory Diseases
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• v.56 no.3
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• pp.248-260
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• 2004

Background : As an effective regimen for isoniazid (INH)-resistant pulmonary tuberculosis, several treatment regimens have been recommended by many experts. In Korea, a standard regimen has not been established for INH-resistant tuberculosis, and the treatment by individual physicians has been performed on an empirical bases. The purpose of the present study was to retrospectively describe the treatment characteristics and evaluate the treatment outcomes of patients with INH-resistant tuberculosis. Materials and Methods : Sixty of 69 patients reported to have INH-resistant tuberculosis from 1994 to 2001 were retrospectively analyzed. Exclusion criteria included: death from other causes, with the exceptions of tuberculosis and incomplete treatment, including a patient's transfer-out. Results : A previous tuberculosis history was found in 28 (46.7%) patients. The sputum smear for acid-fast bacilli was positive in 44 (73.3%) patients, and 30 (50.0%) had cavitary disease. Streptomycin resistance coexisted in 25.0% of isolates. INH was to be prescribed continuously, even after INH resistance was reported, in 86.0% of patients. The treatment regimens were diverse between the patients according to drug regimen composition and treatment duration. The most frequent prescribed regimen included rifampin, ethambutol and pyrazinamide, with and without INH, for the full 12-month term of treatment. Treatment failure occurred in 13 (21.7%) patients. Cavitary disease (p=0.005) and a treatment regimen with second-line drugs, excluding rifampin (p=0.015), were associated with treatment failure. One patient experienced a relapse. Conclusions : Standardized treatment guidelines will be needed in Korea to improve the treatment efficacy for INH-resistant tuberculosis.

• Tuberculosis and Respiratory Diseases
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• v.69 no.2
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• pp.95-102
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• 2010

Background: The increasing rate of drug-resistant tuberculosis (TB) is a threat to the public health and TB control. In Korea, about 75~80% of TB patients are treated in private hospitals and the rate has been continuously increasing since 2000. Methods: On a retrospective basis, we enrolled 170 newly diagnosed with or retreated for multidrug-resistant TB (MDR-TB) in 2004 from 21 private hospitals. We extracted the following demographics and treatment history from patient medical records: initial treatment outcomes, cumulative survival rates, treatment outcomes, and prognostic factors. Results: Of the 170 patients, the majority were male (64.1%), the mean age was 44.5 years old, and mean body-mass-index was $20.2kg/m^2$. None of the patients tested positive for HIV. Eleven (6.5%) were confirmed to have extensively drug-resistant TB (XDR-TB) at treatment initiation. Treatment success rates were not different between XDR-TB (36.4%, 4/11) and non-XDR MDR-TB (51.6%, 82/159). Default rate was high, 21.8% (37/170). Far advanced disease on X-ray was a significant negative predictor of treatment success; advanced disease and low BMI were risk factors for all-cause mortality. Conclusion: In private hospitals in Korea, the proportion of XDR-TB in MDR-TB was comparable to previous data. The treatment success rate of MDR-/XDR-TB remains poor and the failure rate was quite high. Adequate TB control policies should be strengthened to prevent the further development and spread of MDR-/XDR-TB in Korea.

• Journal of Preventive Medicine and Public Health
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• v.16 no.1
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• pp.41-50
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• 1983

The incidence of sexually transmitted disease, especially gonorrhoea has risen despite the progress in its diagnosis and treatment. For the effective control programs of sexually transmitted disease, it should be required socio-medical approaches. A study on gonorrhoea and penicillinase-producing Neisseria gonorrhoeae(PPNG) was conducted in Jeonju and Kunsan area from March, 1982 through August, 1982. The 221 entertrainers were studied in order to determine the prevalences of gonorhoea and PPNG. Socio-demographic informations of the entertainers were obtained by interviewing them. Gonococci were cultured on Thayer-Martin enrichment media for isolation, and PPNG was confirmed using beta-lactamase reagent ($PADAC^{tm}$ Beta-Lactamase Test Strips, Galbiochem-Behring). The results of the study are summarized as follows; 1. The average age of the entertainers studied was $26.1{\pm}4.7$ years. 2. The average years of working in entertaining business was $2.4{\pm}1.4$ years, and the average income per month was $239,592{\pm}90,480$ won. On the education level, 70.6% of the entertainers were middle or high school graduates 3. 47.5% of the entertainers were using contraceptives. 90.5% have experienced artificial abortion. 4. 37(16.7%) out of 221 entertainers were revealed to gonorrhoea, and 13 (35.1%) of gonorrhoea patients were by PPNG. 5. The prevalence rates of gonorrhoea and the proportion of PPNG by age were not significant statistically. Meanwhile, the colelations between the rates of gonorrhoea and education, frequency of love-making with customers and type of sexual partner were highly significant statistically. 6. 37 strains of gonococci isolated were almostly resistant to several antimicrobial agents, especially amikacin, clindamycin and chloramphenicol. Furthermore PPNG strains were completely resistant to not only above drugs but also penicillin.

• Journal of Life Science
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• v.17 no.3 s.83
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• pp.386-395
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• 2007

A total of 27 strains of Vibrio parahaemolyticus (18 strains isolated from Korea and 9 strains from Japan) were serotyped and examined for biochemical characteristics, antimicrobial susceptibility patterns, cytotoxicity assay, thermostable direct hemolysin (TDH) production and molecular epidemiology. Using polymerase chain reaction (PCR) method and DNA probe hybridization method, the strains were tested for toxR, tdh, trh and ORF 8 genes. The V. parahaemolyticus isolated from patients were belonged to 8 different serotypes : O3:K6, O1:K38, O3:K57, O4:K9, O4:Kl2, O4:K68, O5:Kl5 and O6:K46. Urease-positive strain possessed the trh gene, and conversely, urease-negative strains lacked the gene, indicating that urease production by V. parahemolyticus strains strongly correlates with the possession of the trh gene. Most strains showed multiple resistant to more than three antibiotics and the antibiogram could be classified into 6 group (I to VI). All of the O3:K6 strains isolated in South Korea and Japan producted TDH at high levels. The TDH titers ranged between 256 and 2.048, and the average titer was 1009. To distinguish the new and increasingly common V. parahaemolyticus strains from clinical isolates, ORF 8 is a useful genetic marker. After Southern hybridization, the HindIII restriction fragment patterns of the tdh gene were grouped one type, respectively. One type showed two bands one of which was 4.3kb and the other was 11.5kb in size. Variation between the O3:K6 serotype are minor when compared to the differences seen with the non O3:K6 strains. The migration patterns of Not I -digested of the total DNA of the O3:K6 strains were similar, and only slight variations were observed between the serotypes. By contrast, the O3:K6 strains and non O3:K6 had markedly different profiles. In conclusion, Random amplified polymorphic DNA (RAPD) profile using appropriate primers was an effective epidemiological marker.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.5
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• pp.796-801
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• 2001

A total of 277 mineral spring water samples in Kangwon province from 1999 to 2000 were analyzed for the presence of Yersinia spp. by the conventional Food and Drug Administration protocol, and presumptive strains were identified by morphological, cultural and biochemical tests according to Bergey’s manual. Also, the biotypes, serotypes, and susceptibility to 12 antibiotics were tested. Among the total 277 mineral spring water samples, 40 samples (14.4%) were found to be contaminated with Yersinia species. Among the 40 strains of Yersinia spp. isolates, 33 strains (82.5%) for Yersinia enterocolitica, 4 strains (10%) for Yersinia frederiksenii, 2 strains (5%) for Yersinia intermedia, and 1 strain (2.5%) for Yersinia sakazaki were identified, respectively. Of 40 Yersinia spp. isolates, Yersinia enterocolitica (82.5%) was the most predominant species in the mineral spring water samples compared to other Yersinia species. Compared to direct culture method after KOH treatment and KOH treatment method after cold enrichment for better isolation ratio of according to comparision of Yersinia species, the detection ration (18.5%) of KOH treatment method after cold enrichment was about 3 times better than that (6.1%) of direct culture method after KOH treatment. According to serotypes of Y. enterocolitica isolates, O : 5 (12.9%) was the most predominant and followed by O : 3 (9.7%), O : 8 (6.5%), and O : 9 (3.2%), and others. For biotypes of Y. enterocolitica isolates, 1A (71.0%) was the most predominantly abundant and followed by 3A (12.9%), 3B (9.7%), 1B (3.2%) and 5 (3.2%). Also, an antibiotic susceptibility test showed that Yersinia spp. isolates were very susceptible to the antibiotics tested, but they were very strongly resistant to ampicillin, cephalothin and carbenicillin.

• Korean Journal of Clinical Laboratory Science
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• v.53 no.3
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• pp.225-232
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• 2021

This study sought to investigate the distribution, antimicrobial resistance rate, and bacterial co-infection frequency of non-tuberculous mycobacteria (NTM) in a single center in Incheon, South Korea. A total of 8,258 specimens submitted for tuberculosis (TB)/NTM real-time PCR tests during the years 2015 to 2020 were retrospectively reviewed. In total, 296 specimens (3.6%) were NTM positive, and the positivity increased from 2.5% (30/1,209) in 2015 to 3.8% (66/1,740) in 2020. Of 296 NTM specimens, 54.7% (162/296) were identified as the Mycobacterium avium complex (MAC) followed by the Mycobacterium abscessus complex (MABC) 20.9% (62/296), M. fortuitum 6.4% (19/296) and M. flavescens 3.4% (10/296). Of the NTM-positive specimens, 76.7% (227/296) were tested for drug resistance. The results showed multidrug-resistant NTM in 40.1% (91/227) and extensively drug-resistant NTM in 59.9% (136/227) of these specimens. Of the 145 isolates taken for bacterial culture, bacteria/fungi co-infection with NTM accounted for 43.4% (63/145), in which the most common bacterial species was Klebsiella pneumonia (23.8%, 15/63). This study is the first report on the distribution and antimicrobial resistance of NTM in Incheon. As the proportion of NTM infections increases, active treatment and thorough infection control are required for effective management.

• The Journal of the Korean Society for Microbiology
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• v.7 no.1
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• pp.9-15
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• 1972

The authors identified 45 Shigella cultures among 63 suspectable cultures obtained from the cases showing dysentery or the like symptoms in various parts of the country during the period from February to September, 1971. Of 45 cultures, 36 cultures belonged to Subgroup B, 7 cultures to Subgroup C and 2 cultures to Subgroup D. There was none of cultures belonging to Subgroup A in 1971, although the authors detected one culture of that among 41 identified Shigella in 1967. Of 36 cultures belonging to Subgroup B, 27 cultures were $B_{2a}$ and 3 cultures of each $B_{3c},\;B_{4a}$ and $B_y$ were serotyped, respectively. Of 7 cultures beloning to Subgroup C, one was $C_{11}$ and other six cultures were $C_{15}$ and all S. boydii were isolated in Seoul area. It might be possible to suggest that there was a tendency of decreasing prevalence of S. dysenteriae and of increasing prevalence of S. boydii in Korea, although the number of cultures. tested were very much limited to conclude, if it was compared with the results obtained by the authors in 1967 that the fourty-one identified Shigella were composed of one culture of S. dysenteriae, 36 cultures of S. flexneri, 4 cultures of S. sonnei and none of S. boydii. According to the results obtained from the biochemical tests, the positive ratio of Indol tests in Subgroup B was 31/36, which could be higher than 9/36 observed in 1961, but which was nearly correlated with the result published by Ewing and his colleagues in U.S.A. The positive rates of both sorbitol and raffinose were lower than that observed by Ewing and his colleagues in S. flexneri. Regarding with the sensitivity of Shigella cultures to the antibiotics being widely used in the country, 46.2% were sensitive to chloramphenicol, which was very much higher than that observed by Park, and 85.8% were sensitive to ampicillin, which would be the drug of choice according to the results from the In Vitro tests.

• Current Research on Agriculture and Life Sciences
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• v.4
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• pp.132-140
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• 1986

Eighty one products from 36 kinds of vitamin and mineral feed supplement collected during August, 1984 to February, 1985 were examined for microbiological contamination. In addition, 83 strains of coliform isolated from the samples were tested for the resistance to 8 kinds of antimicrobial drugs and distribution of R plasmid. General bacteria were detected in all of samples tested. Bacterial population was varied from less than 10 per gram of the sample to 1,400,000 per gram and 34 (42%) of 81 samples were contaminated with 100 to 1,000 cells per gram. Coliform isolation, which was more frequent in samples with larger number of general bacteria, was possible in 14 (17.3%) out of 81 samples tested and 6 (33.3%) out of 18 companies were coliform positive in their products. Forty one (49.4%) out of 83 coliform isolates were fecal coliform. The frequency of resistant strains was the highest to sulfadimethoxine (Sa) with 92.8% and followed by streptomycin (Sm, 67.5%), tetracycline (Tc, 50.6%), kanamycin (Km, 26.5%), chloramphenicol (Cm, 18.1%) and ampicillin (Am, 15.7%). No strain was resistant to nalidixic acid (Na) and gentamicin (Gm). The resistance frequency of fecal coliform strains were higher compare to non-fecal coliform strains. There were minimum inhibitory concentration (MIC) of $3,200{\mu}g/m{\ell}$ or higher in 7 strains to Am, 3 to Sm and 3 to Km, and 70 strains had MIC of $1,600{\mu}g/m{\ell}$ of higher to Sa while Tc had MICs from $1.6{\mu}g/m{\ell}$ to $400{\mu}g/m{\ell}$. All strains had MICs of $6.3{\mu}g/m{\ell}$ of lower to Na and $3.1{\mu}g/m{\ell}$ of lower to Gm. Seventy nine (95.2%) of 83 strains were resistant to one or more drugs tested. The most frequent resistance patterns were SaSm (14.5%) and followed by SaSmTc(12%), SaSmTcKm(8.4%) SaTc (8.4%) and SaSmKm (7.2%) ; total 19 different patterns were noted. Thirty two (40.5%) of 79 resistant strains were transferred all of a part of their resistance to Escherichia coli ML 1410. The frequency of transferable resistance was high in Am (100%) and Cm (80%) while low in Tc (38.1%), Sa (18.2%), Sm (17.9%) and Km (4.5%).