• Title/Summary/Keyword: mycobacterium

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Differentiation of four Mycobacterium Species using DNA-DNA Hybridization Method using Specific Probes

  • Kweon, Tae-Dong;Bai, Sun-Joon;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2013.05a
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    • pp.1012-1014
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    • 2013
  • DNA-DNA hybridization method with four oligonucleotide-specific probes was used simultaneously for differentiation and identification of four Mycobacterium species (Mycobacterium tuberculosis, M. avium, M. intracellulare, and M. kansasii). This DNA-DNA hybridization method with 4 oligonucleotide-specific probes, which targets in the rpoB region of 4 Mycobacteria species, respectively, was tested on 322 clinical isolates. Using DNA-DNA hybridization method, we detected M. tuberculosis (282 strains), M. avim (7 strains), M. intracellulare (9 strains), and M. kansasii (3 strain) from 322 clinical isolates. This result was compared with conventional biochemical test and rpoB DNA sequence analysis of this clinical isolates. We confirmed identification of Mycobacterium tuberculosis, M. avium, M. intracellulare, and M. kansasii with high sensitivity (100 %) and specificity (100 %). This DNA-DNA hybridization method could be performed within 4 hours at least. Therefore, we suggest that DNA- DNA hybridization method using 4 rpoB DNA probes of Mycobacteria could be used for accurate, rapid, convenient detection and identification of Mycobacterium tuberculosis, M. avium, M. intracellulare, and M. kansasii in clinical samples.

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Mixed Infection of Mycobacterium abscessus subsp. abscessus and Mycobacterium tuberculosis in the Lung

  • Sohn, Sungmin;Wang, Sungho;Shi, Hyejin;Park, Sungrock;Lee, Sangki;Park, Kyoung Taek
    • Journal of Chest Surgery
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    • v.50 no.1
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    • pp.50-53
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    • 2017
  • A mixed infection of Mycobacterium abscessus subsp. abscessus (Mab) and Mycobacterium tuberculosis (MTB) in the lung is an unusual clinical manifestation and has not yet been reported. A 61-year-old woman had been treated for Mab lung disease and concomitant pneumonia, and was diagnosed with pulmonary tuberculosis (PTB). Despite both anti-PTB and anti-Mab therapy, her entire left lung was destroyed and collapsed. She underwent left pneumonectomy and received medical therapy. We were able to successfully treat her mixed infection by pneumonectomy followed by inhaled amikacin therapy. To the best of our knowledge, thus far, this is the first description of a mixed Mab and MTB lung infection.

Studies on Identification and Drug Resistance of Atypical Mycobacteria isolated from Patients with Pulmonary Tuberculosis (폐결핵환자에서 비정형항산균의 분리, 동정 및 약제감수성에 관하여)

  • Chung, Dong-Hyun;Kim, Sung-Kwang;Kim, Joo-Deuk
    • Journal of Yeungnam Medical Science
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    • v.1 no.1
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    • pp.49-58
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    • 1984
  • The differential diagnosis of atypical mycobacteriosis caused by atypical mycobacteria (with the exception of Mycobacterium tuberculosis, Mycobacterium bovis, and Mycobacterium leprae) which are widly distributed in soil and water, from pulmonary tuberculosis is possible only when atypical mycobacteria are isolated and identified. In this investigation, attempts were made to isolate atypical mycobacteria from persons registered as tuberculosis patients in the Anyang Health Center in Anyang City, Kyungki province, Korea. Biological and biochemical tests were performed for the atypical mycobacteria isolated from these patients, also retrospective analysis of clinical and X-ray findings of the patients with bacteriologically confirmed atypical mycobacteriosis were done. The results can be summarized as follows: 1. 103 strains of mycobacteria were isolated among 334 sputum samples from patients. 2. Among the isolated mycobacteria, 10 strains (9.7%) were found to be a atypical mycobacteria and 93 strains (90.3%) were tubercle bacilli of human type. 3. On the basis of Runyon's grouping of atypical mycobacteria, there were 3 strains (30.0 %) of scotochromogen and nonphotochromogen respectively, 4 strains (40.0%) of rapid grower, and no photochromogen. 4. By biochemical tests, 3 strains of scotochromogen were identified as Mycobacterium scrofulaceum (2 strains) and Mycobacterium szulgai (1 strain) 3 strains of nonphotochromogen were Mycobacterium avium-complex (2 strains) and Mycobacterium terriae (1 strain), and 4 strains of rapid grower were Mycobacterium fortuitum (3 strains) and Mycobacterium chelonei. 5. In drug sensitivity tests, all 10 strains isolated atypical mycobacteria showed resistance to various concentration of INH and SM and low concentration (10mcg, 40mcg and 50mcg) of EB, TH, and CS, and were sensitive to only high concentration (20mcg and 100mcg) of EB, TH, CS, and RFP. 6. In analysis of clical findings by the patients with bacteriologically confirmed atypical mycobacteriosis, it was found that clinical symptoms of these patients appeared not to be mild than those of patients with pulmonary tuberculosis. The patients with atypical mycobacteriosis had been treated for pulmonary tuberculosis for a long time and they showed no improvement.

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Application of Polymerase Chain Reaction (PCR) to the Diagnosis of Tuberculosis (Polymerase Chain Reaction (PCR)을 이용한 결핵의 진단에 관한 연구)

  • Kim, Ho-Joong;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Kim, Keun-Youl;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.6
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    • pp.517-525
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    • 1992
  • Background: Since its development by Saiki et al, polymerase chain reaction (PCR) has been very useful in various fields of molecular biology. PCR can be used for the detection of a very small amount of microbial agent, and is especially useful in those patients who are difficult to diagnose microbiologically or serologically. Mycobacterium tuberculosis is a very slowly growing organism and AFB staining frequently shows false negative results, and therefore PCR would be a very rapid, easy, and sensitive diagnostic method for the diagnosis of Mycobacterium tuberculosis. Method: To compare PCR with conventional methods in diagnosing Mycobacterium tuberculosis in sputum, we used sputa of patients who visited or were admitted to Seoul National University Hospital. The amplification targets were 383 base pair DNA, a part of 2520 base pair DNA encoding 65 kD Mycobacterium tuberculosis specific protein (the primers are TB-1, -2), and 123 base pair DNA, a part of IS6110 fragment, which multiple copies are known to exsist PCR one genome (the primers are Sal I-1, -2). We also requested AFB staing and culture to the lab of Seoul National University Hospital with the same sample and compared the results. Results: 1) Using TB-1, -2 primers, PCR was positive in 73.1% (19/26) of culture positive sputa, in 12.5% (1/8) of culture negative. but clinically diagnosed tuberculous sputa, and was negative in all sputa of patients who were clinically diagnosed as non-tuberculous etiology. 2) Using Sal I-I, -2 primers, PCR was positive in 94.1% (32/34) of culture positive sputa, in 23.1% (6/26) of culture negative, but clinically diagnosed tuberculous sputa, and was negative in 87.5% (14/16) of sputa from patients who were clinically diagnosed as non-tuberculous etiology. Conclusion: PCR could be a very rapid, sensitive and specific method for the diagnosis of Mycobacterium tuberculosis in sputa, and further studies should be followed for the development of easier method.

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Development of New Vector Systems as Genetic Tools Applicable to Mycobacteria (Mycobacteria에 적용 가능한 genetic tool로서의 새로운 vector system 개발)

  • Jeong, Ji-A;Lee, Ha-Na;Ko, In-Jeong;Oh, Jeong-Il
    • Journal of Life Science
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    • v.23 no.2
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    • pp.290-298
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    • 2013
  • The genus Mycobacterium includes crucial animal and human pathogens such as Mycobacterium tuberculosis, Mycobacterium leprae, and Mycobacterium bovis. Although it is important to understand the genetic basis for their virulence and persistence in host, genetic analysis in mycobacteria was hampered by a lack of sufficient genetic tools. Therefore, many functional vectors as molecular genetic tools have been designed for understanding mycobacterial biology, and the application of these tools to mycobacteria has accelerated the study of mechanisms involved in virulence and gene expression. To overcome the pre-existing problems in genetic manipulation of mycobacteria, this paper reports new vector systems as effective genetic tools in Mycobacterium smegmatis. Three vectors were developed; pKOTs is a suicide vector for mutagenesis containing a temperature-sensitive replication origin (TSRO) and the sacB gene encoding levansucrase as a counterselectable marker. pMV306lacZ is an integrative lacZ transcriptional fusion vector that can be inserted into chromosomal DNA by site-specific recombination. pTnMod-OKmTs is a minitransposon vector harboring the TSRO that can be used in random mutagenesis. It was demonstrated in this study that these vectors effectively worked in M. smegmatis. The vector systems reported here are expected to successfully applicable to future research of mycobacterial molecular genetics.

Study on the Growth Factors for Rapidly Cultivating Mycobacterium spp. (마이코박테리움을 신속하게 배양할 수 있는 성장 인자에 관한 연구)

  • Ha, Sung-Il;Park, Kang-Gyun;Suk, Hyun-Soo;Shin, Jeong-Seob;Shin, Dong-Pil;Kwon, Min-O;Park, Yeon-Joon
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.2
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    • pp.177-184
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    • 2019
  • Mycobacteria grow slowly. Therefore, a solid medium should be used for eight weeks and a liquid medium for six weeks. The purpose of this study was to find the growth factors that can grow Mycobacterium rapidly and to help develop a solid medium for rapid identification. Three types of Mycobacterium growth factors were evaluated with 10 Mycobacteria by adding activated charcoal, defibrinated sheep blood, and L-ascorbic acid to $Difco^{TM}$ Mycobacteria 7H11 agar (Becton, Dickinson and Company, Sparks, MD, USA). The time to detection and the distinguishability of a colony were compared with that of the current method. In the rapidly growing Mycobacterium, the difference in detection time between the new media and conventional media confirmed that the new media was faster. M. kansasii and M. intracelluare grew faster in 7H11 C than in 7H11 medium. MTB grew faster than the other media in 7H11 C. This study confirmed that the two growth factors affect fast-growing Mycobacteria and slow-growing Mycobacteria. 7H11 C showed better distinguishability than the conventional media in all 10 Mycobacterium due to the color contrast. In particular, when the MTB was grown, the size of the colonies was larger than with other media, so visualization was easy.

A Case of Exit-Site Infection and Abscess by $Mycobacterium$ $abscessus$ in a CAPD Patient (지속적외래복막투석 환자에서 발생한 $Mycobacterium$ $abscessus$에 의한 출구 감염 및 농양 1예)

  • Jung, Sun-Young;Na, Ji-Hoon;Cho, Kyu-Hyang;Park, Jong-Won;Do, Jun-Young;Yun, Kyeung-Woo;Song, In-Wook;Cho, Jeong-Hwan;Son, Chang-Woo
    • Journal of Yeungnam Medical Science
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    • v.26 no.2
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    • pp.137-142
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    • 2009
  • Nontuberculous mycobacterial infections are a rare, but clinically important cause of infections in continuous ambulatory peritoneal dialysis (CAPD) patients. This is typically suspected when a patient does not respond to treatment with the usual antibiotics. We describe here a case of $Mycobacterium$ $abscessus$ exit site infection with abdominal wall abscess formation that was associated with CAPD, which required peritoneal catheter removal, surgical debridement of the abscess and long term antibiotic therapy.

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Non-Tuberculous Mycobacterium Induced Pseudoaneurysm of the Common Carotid Artery

  • Lee, Hae Young;Cho, Seong Ho;Kim, Hyun Su;Moon, Jeong Min;Lee, Sangho;Kim, Jong In
    • Journal of Chest Surgery
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    • v.49 no.6
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    • pp.468-471
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    • 2016
  • An 81-year-old male patient presented with complaint of a pulsating neck mass. The patient had a previous history of cervical lymphadenopathy by non-tuberculous mycobacterium infection. Rapid growth of the mass on admission and contrast enhanced computed tomography of the neck resulted in a diagnosis of non-tuberculous mycobacterium induced pseudoaneurysm. The patient underwent emergency open repair of the pseudoaneurysm. Pseudoaneurysm of the common carotid artery is regularly reported, but here we report a rare case of non-tuberculous mycobacterium induced pseudoaneurysm of the common carotid artery.

Inhibition of the DevSR Two-Component System by Overexpression of Mycobacterium tuberculosis PknB in Mycobacterium smegmatis

  • Bae, Hyun-Jung;Lee, Ha-Na;Baek, Mi-Na;Park, Eun-Jin;Eom, Chi-Yong;Ko, In-Jeong;Kang, Ho-Young;Oh, Jeong-Il
    • Molecules and Cells
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    • v.40 no.9
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    • pp.632-642
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    • 2017
  • The DevSR (DosSR) two-component system, which is a major regulatory system involved in oxygen sensing in mycobacteria, plays an important role in hypoxic induction of many genes in mycobacteria. We demonstrated that overexpression of the kinase domain of Mycobacterium tuberculosis (Mtb) PknB inhibited transcriptional activity of the DevR response regulator in Mycobacterium smegmatis and that this inhibitory effect was exerted through phosphorylation of DevR on Thr180 within its DNA-binding domain. Moreover, the purified kinase domain of Mtb PknB significantly phosphorylated RegX3, NarL, KdpE, TrcR, DosR, and MtrA response regulators of Mtb that contain the Thr residues corresponding to Thr180 of DevR in their DNA-binding domains, implying that transcriptional activities of these response regulators might also be inhibited when the kinase domain of PknB is overexpressed.

An Unusual Case of Post-Operative Spondylitis Caused by $Mycobacterium$ $Intracellulare$ in an Immunosuppressed Patient

  • Kim, Sung-Hoon;Son, Dong-Wuk;Lee, Sang-Weon;Song, Geun-Sung
    • Journal of Korean Neurosurgical Society
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    • v.50 no.5
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    • pp.460-463
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    • 2011
  • There are few reported cases of post-operative spondylitis caused by $Mycobacterium$ $Intracellulare$. A 75-year-old female presented to our hospital with low back pain and paraparesis after a fall. The radiologic examination revealed compression fractures of L1, L3 and L4 and an epidural hematoma compressing the spinal cord. The dark-red epidural hematoma was urgently evacuated. Four weeks post-operatively, neurologic deficits recurred with fever. On magnetic resonance image, an epidural abscess and osteomyelitis were detected in the previous operative site. Five weeks post-operatively, revision was performed with multiple biopsies. The specimen were positive for acid-fast bacilli and traditional anti-tuberculous medications were started. Because the Polymerase Chain Reaction for non-tuberculous mycobacterium (NTM) was positive, the anti-tuberculous medications were changed to anti-NTM drugs. However, the neurologic deficits did not improve and persistent elevation of erythrocyte sedimentation rate and C-reactive protein were noted. Eight weeks after the revision, $Mycobacterium$ $Intracellulare$ was detected in the specimen cultures. Despite supportive care with medication, the patient died due to multiple organ failure.