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Trial for Drug Susceptibility Testing of Mycobacterium tuberculosis with Live and Dead Cell Differentiation (세포 염색 방법을 이용한 결핵균 감수성 검사법)

  • Ryu, Sung-Weon;Kim, Hyun-Ho;Bang, Mun-Nam;Park, Young-Kil;Park, Sue-Nie;Shim, Young-Soo;Kang, Seongman;Bai, Gill-Han
    • 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.

A Study on the Trend and Utilization of Stone Waste (석재폐기물 현황 및 활용 연구)

  • Chea, Kwang-Seok;Lee, Young Geun;Koo, Namin;Yang, Hee Moon
    • Korean Journal of Mineralogy and Petrology
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    • v.35 no.3
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    • pp.333-344
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    • 2022
  • The quarrying and utilization of natural building stones such as granite and marble are rapidly emerging in developing countries. A huge amount of wastes is being generated during the processing, cutting and sizing of these stones to make them useable. These wastes are disposed of in the open environment and the toxic nature of these wastes negatively affects the environment and human health. The growth trend in the world stone industry was confirmed in output for 2019, increasing more than one percent and reaching a new peak of some 155 million tons, excluding quarry discards. Per-capita stone use rose to 268 square meters per thousand persons (m2/1,000 inh), from 266 the previous year and 177 in 2001. However, we have to take into consideration that the world's gross quarrying production was about 316 million tons (100%) in 2019; about 53% of that amount, however, is regarded as quarrying waste. With regards to the stone processing stage, we have noticed that the world production has reached 91.15 million tons (29%), and consequently this means that 63.35 million tons of stone-processing scraps is produced. Therefore, we can say that, on a global level, if the quantity of material extracted in the quarry is 100%, the total percentage of waste is about 71%. This raises a substantial problem from the environmental, economical and social point of view. There are essentially three ways of dealing with inorganic waste, namely, reuse, recycling, or disposal in landfills. Reuse and recycling are the preferred waste management methods that consider environmental sustainability and the opportunity to generate important economic returns. Although there are many possible applications for stone waste, they can be summarized into three main general applications, namely, fillers for binders, ceramic formulations, and environmental applications. The use of residual sludge for substrate production seems to be highly promising: the substrate can be used for quarry rehabilitation and in the rehabilitation of industrial sites. This new product (artificial soil) could be included in the list of the materials to use in addition to topsoil for civil works, railway embankments roundabouts and stone sludge wastes could be used for the neutralization of acidic soil to increase the yield. Stone waste is also possible to find several examples of studies for the recovery of mineral residues, including the extraction of metallic elements, and mineral components, the production of construction raw materials, power generation, building materials, and gas and water treatment.

A Color-Reaction-Based Biochip Detection Assay for RIF and INH Resistance of Clinical Mycobacterial Specimens

  • Xue, Wenfei;Peng, Jingfu;Yu, Xiaoli;Zhang, Shulin;Zhou, Boping;Jiang, Danqing;Chen, Jianbo;Ding, Bingbing;Zhu, Bin;Li, Yao
    • Journal of Microbiology and Biotechnology
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    • v.26 no.1
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    • pp.180-189
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    • 2016
  • The widespread occurrence of drug-resistant Mycobacterium tuberculosis places importance on the detection of TB (tuberculosis) drug susceptibility. Conventional drug susceptibility testing (DST) is a lengthy process. We developed a rapid enzymatic color-reaction-based biochip assay. The process included asymmetric multiplex PCR/templex PCR, biochip hybridization, and an enzymatic color reaction, with specific software for data operating. Templex PCR (tem-PCR) was applied to avoid interference between different primers in conventional multiplex-PCR. We applied this assay to 276 clinical specimens (including 27 sputum, 4 alveolar lavage fluid, 2 pleural effusion, and 243 culture isolate specimens; 40 of the 276 were non-tuberculosis mycobacteria specimens and 236 were M. tuberculosis specimens). The testing process took 4.5 h. A sensitivity of 50 copies per PCR was achieved, while the sensitivity was 500 copies per PCR when tem-PCR was used. Allele sequences could be detected in mixed samples at a proportion of 10%. Detection results showed a concordance rate of 97.46% (230/236) in rifampicin resistance detection (sensitivity 95.40%, specificity 98.66%) and 96.19% (227/236) in isoniazid (sensitivity 93.59%, specificity 97.47%) detection with those of DST assay. Concordance rates of testing results for sputum, alveolar lavage fluid, and pleural effusion specimens were 100%. The assay provides a potential choice for TB diagnosis and treatment.

Pulmonary Resection Combined with Isoniazid-and Rifampin-based Drug Therapy for Patients with Multidrug-resistant Tuberculosis (다제내성 폐결핵 환자에서 폐절제술 후 일차 항결핵제 치료)

  • Park, Seung-Kyu;Kim, Jin-Hee;Kim, Jun-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.59 no.2
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    • pp.179-185
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    • 2005
  • Background : To evaluate the clinical efficacy of pulmonary resection combined with first-line antituberculous drug therapy in patients with well-localized, cavities-containing pulmonary multidrug-resistant tuberculosis (MDR-TB). Method : From February 1998, seventeen patients with well-localized, cavities-containing pulmonary MDR-TB were enrolled and followed prospectively up to December 2004. After radical pulmonary resection, the patients were treated with antituberculous drugs comprising of isoniazid (H), rifampin (R), pyrazinamide (Z), ethambutol (E), and streptomycin (S) (3HERZS/3HERS/6HER). Results : All recovered isolates of M. tuberculosis were resistant to both isoniazid and rifampin, and to a mean of 4.8 antituberculous drugs (range, 2 to 7 drugs). Surgical procedures included lobectomy (13 patients), lobectomy plus segmentectomy (3 patients), and pneumonectomy (1 patient). The median time for postoperative sputum smear and culture conversion was 2 days (range, 1 to 23 days). Fifteen (94%) patients had durable cures (mean follow-up period, 39.0 months). One patient failed to convert her sputum and was successfully switched to second-line therapy; one patient developed active disease again almost 7 years later, likely due to re-infection with a new M. tuberculosis strain. Conclusion : Radical resection combined with administration of first-line antituberculous agents was effective in patients with well-localized, cavities-containing pulmonary MDR-TB.

A Clinical Effect of Retreatment by Prothionamide, Cycloserine, Para-Aminosalicylic Acid, Streptomycin (Kanamycin or Tuberactinomycin) on Pulmonary Tuberculosis (폐결핵 재치료의 Prothionamide, Cycloserine, Para-Aminosalicylic Acid, Streptomycin (Kanamycin or Tuberactinomycin) 4제 요법 임상 효과)

  • Shin, Cheol-Shick;Im, Young-Jae;Kim, Young-Jun;Koh, Seok-Shin;Kim, Moon-Shik
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.2
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    • pp.167-171
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    • 1992
  • Background:In the management of patients whose primary chemotherapy has failed, very careful assessment is essential. It is important to find out as accurate a chemotherapy history as possible. Preferably it should contain the drugs which has never used before. The present report concerns the results of retreatment of pulmonary tuberculosis patients treated at National Kongju Tuberculosis Hospital. Method: A retrospective study was made through the regular follow-up of 112 smear positive cases, who were treated by four-drug regimen between July 1985 and June 1990. Four drugs were, namely prothionamide, cycloserine, para-aminosalicylic acid, and streptomycin (kanamycin or tuber-actinomycin). The duration of follow-up was over one year. Results: 1) Out of 112 cases with positive sputum AFB smear, 72 (64%) achieved the negative conversion. 2) Among the 72 patients, 85% achieved negative conversion within 3 months after treatment. 3) When the duration of patient's illness was less than 2 years, 2 to 4 years and more than 5 years, the favourable response to retreatment was 86%, 62% and 54%, respectively. 4) When the number of sensitive drugs was 4,3,2 and 1, the favourable response rate was 74%, 68%, 39% and 0%, respectively. Conclusion: The shorter the duration of patient's illness was, the larger the number of sensitive drugs was. And the larger the number of sensitive drugs was, the better the result of treatment was. Thus it is very crucial to successfully treat newly discovered patients with adequate regimens and proper case-holding.

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The Prevalence and Risk Factors of Drug Resistant Pulmonary Tuberculosis Investigated at One University Hospital in Seoul (서울지역 한 대학병원에서 조사된 폐결핵 약제 내성률 및 위험인자)

  • Kim, Do Kyun;Kim, Mi Ok;Kim, Tae-Hyung;Sohn, Jang Won;Yoon, Ho Joo;Shin, Dong Ho;Park, Sung Soo
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.3
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    • pp.243-247
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    • 2005
  • Background : The prevalence of tuberculosis is slowly decreasing in Korea. However, the drug-resistance of pulmonary tuberculosis is a major risk factor of treatment failure. Moreover, the National Surveillance System has recently been discontinued. Therefore, a continuous survey is necessary for the exact detection of the rate of drug resistance. We studied the recent 4-year drug resistance rate of tuberculosis at a single University hospital in Seoul. Materials and Methods : The study included 239 pulmonary tuberculosis patients performed with a tuberculosis culture and a drug-sensitivity test at Hanyang University Medical Center from March 1999 to March 2003. Results : Of the 239 patients included in the study during the 4-year period, 52 patients showed resistance to one or more anti-tuberculosis drug (21.8%). The rate of multi-drug resistance was 12.6%. The resistance rates to isoniazid, rifampin, ethambutol, streptomycin and pyrazinamide were 18.4%, 13.8%, 11.7%, 6.7% and 8.4%, respectively. Ninety patients had a history of previous anti-tuberculosis treatment, and the rates of the overall drug resistance and multi-drug resistance of these patients were 36.7% and 25.6%, respectively. The patients with drug-resistance showed a higher rate of a previous tuberculosis treatment history (63.5%) than the drug-sensitive group patients (30.5%). Conclusion : The rate of drug resistant tuberculosis is 21.8%, and multi-drug resistant tuberculosis is 12.6%. The rate of drug resistance is higher in those previously treated for tuberculosis.

Evaluation of Liquid Culture System in Sputum Culture and Drug Susceptibility of Mycobacterium tuberculosis (고위험병원체 결핵균의 신속진단을 위한 액체배양시스템 평가)

  • Kim, Jin-Sook;Kim, Seung-Cheol;Jeon, Bo-Young;Park, Seung-Kyu
    • Korean Journal of Microbiology
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    • v.45 no.3
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    • pp.281-285
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    • 2009
  • In this study, we compared the BacT/Alert liquid culture system with Ogawa and $L\ddot{o}wenstein$-Jensen (L-J) media for sputum culture and drug susceptibility test (DST) of Mycobacterium tuberculosis. Rapid liquid culture systems have been widely employed both for primary cultures of M. tuberculosis from clinical specimens and for drug susceptibility test because of its greater sensitivity and faster turn-around time than the conventional egg-based culture methods (Ogawa, $L\ddot{o}wenstein$-Jensen media). Sputum specimens were decontaminated with N-acetyl-L-cysteine (NALC)-4% NaOH and inoculated into the BacT/Alert culture bottles and Ogawa media. 95 from among 135 sputa were smear-positive, 97 (71.9%) were culture-positive by the BacT/Alert culture system, while 89 (65.9%) were positive by Ogawa media. The mean time to culture-positive by the BacT/Alert process system was about 11.3 days, which was significantly shorter than that by Ogawa media (22.4 days). Of 32 M. tuberculosis cultures examined for drug sensitivity, the concordant rate between the two methods (BacT/ Alert liquid culture system, $L\ddot{o}wenstein$-Jensen media) ranged from 87.5% for isoniazid and 90.6% for rifampicin.

Outcome of Chemotherapy with Pulmonary Tuberculosis Resistant to Isoniazid and Rifampin (다제내성 폐결핵의 화학치료)

  • Park, Seung-Kyu;Kwon, Eun-Soo;Ha, Hyun-Cheol;Hwang, Su-Hee
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.1
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    • pp.25-35
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    • 1999
  • Background : The treatment for multi-drug resistant tuberculosis(MDRTB) is encountered to be important clinically, but there are still a few reports about it all over the world. So, we evaluated the outcomes of only chemotherapy for the pulmonary MDRTB retrospectively. Method: We reviewed the clinical courses of 63 patients with pulmonary disease due to M.tuberculosis resistant to rifampin and isoniazid who were under follow-up between March 1996 and June 1996 after hospitalization at our hospital between January 1993 and January 1996. We performed cohort retrospective study for all these patient's records. Their regimens were selected individually and preferably included four medications that they had not been given previously and to which the strain was fully susceptible. Results: The 63 patients(mean age, 43.2 years) had previously received a median 5.1drugs. Fifty two(82.5%) patients responded to chemotherapy(as indicated by negative sputum cultures for at least three consecutive months) ; eleven patients(17.5 %) had no response, as shown by continually positive cultures. In a univariate analysis, an unfavorable response was significantly associated with greater number of resistant drugs before the current courses of therapy(relative risk 21.5 ; 95 percent confidence interval, 1.2-3.0; p<0.05). The mean period of follow-up was seventeen months. There was no relapse suooequently among the patients with responses. There was no death related to tuberculosis. Conclusion: In this report from National Masan Tuberculosis Hospital in Korea, multi-drug resistant pulmonary tuberculosis responded relatively well to carefully selected regimens.

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A Case Report of Three Patients with Nontuberculous Mycobacterial Pulmonary Disease Caused by Mycobacterium kansasii (Mycobacterium kansasii에 의한 비결핵성 마이코박테리아 폐질환 3례)

  • Koh, Won Jung;Kwon, O Jung;Suh, Gee Young;Chung, Man Pyo;Kim, Hojoong;Lee, Nam Yong;Kim, Tae Sung;Lee, Kyung Soo;Park, Eun Mi;Park, Young Kil;Bai, Gill Han
    • Tuberculosis and Respiratory Diseases
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    • v.54 no.4
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    • pp.459-466
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    • 2003
  • Mycobacterium kansasii is the second most common cause of nontuberculous mycobacterial pulmonary disease in Western countries and Japan. The clinical and radiological features of pulmonary disease caused by M. kansasii usually resemble those of pulmonary tuberculosis including cavitary infiltrates with an upper lobe predilection. It is also now apparent that patients with M. kansasii pulmonary disease can present with noncavitary nodular bronchiectatic infiltrates similar to lung diseases of M. avium complex. With rifampin-containing regimens, treatment success rates are almost 100%. Timely diagnosis before the development of extensive disease and effective overall treatment strategies are very important to ensure that patients receive the appropriate medications for a sufficiently long period of time. To our knowledge, there has been no Korean case report of M. kansasii pulmonary disease in the immunocompetent patient until now. We report three cases of M. kansasii pulmonary disease in immunocompetent adult patients.

Study on function evaluation tools for stroke patients (뇌졸중(腦卒中) 환자(患者)의 기능평가방법(機能評價方法)에 대(對)한 연구(硏究))

  • Ko, Seong-Gyu;Ko, Chang-Nam;Chox, Ki-Ho;Kim, Young-Suk;Bae, Hyung-Sup;Lee, Kyung-Sup
    • The Journal of Korean Medicine
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    • v.17 no.1 s.31
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    • pp.48-83
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    • 1996
  • Our conclusions for function evaluation tools of Stroke patients are as follows. 1. Evaluating tools of Activities of Daily Living, Katz Index, Barthel Index, Modified Barthel Index have high validity and reliability because of ease of measuring, high accuracy, consistency, sensitivity and sufficient stastistics, but they mainly measure motor function except sense, mentation, language, and social conception. Therefore cerebrovascular disease and brain injury in trauma patients with lacked acknowledgement and sensation, we are not able to apply these tools. 2. PULSES Profile is a useful scale for measuring the patient's over-all status, upper and lower limb functions, sensory components, excretary functions, and intellectual and emotional adaptabilities. It is recognized as a good, useful tool to evaluate patient's whole function. 3. Motor Assessment Scale was designed to measure the progress of stroke patients. The scale was supplemented with upper arm function items. We believe that the Motor Assessment Scale could be a useful evaluation tool with inter-rater reliability ,test-retest reliability. 4. The existing evaluation tools, Katz Index, Barthel Index, Modified Barthel Index, PULSES Profile, Motor Assessment Scale, mainly measured the rehabilitational motor function of sequela of cerebrovascular patients. On the other hand CNS & INH stroke scale can measure cerebrovascular disease patient's neurologic deficits and over-all stautus, which are recognition ability, speech status, motor function, sensory function, activities of daily living. Those scales have been recognized as useful tools to measure function of cerebrovascular disease patients and have increased in use. 5. Every function evaluation tool was recognized to have some validity and inter-rater, test-retest reliability in items of each evaluation tool and total scores of each evaluation tools, but it is thought that none of these scales have been fully validated and proved reliable. Therefore afterward, the development of a highly reliable rating system may best be accomplished by a careful comparison of several tools, using the same patients and the same observers in order to choose the most reliable items from each. 6. Ideal evaluation tools must have the following conditions; (1) It should show the objective functional statues at the same time. (2) It should be repeated consecutively to know changed function status. (3) It should be easy to observe the treatment program. (4) It should have the same result with another rater to help rater exchange information with treatment team members. (5) It should be practical and simple. (6) The patient should not suffer from the observer.

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