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. CONCLUSION: An oligonucleotide chip was developed for the detection of rpoB gene mutations related to drug-susceptibility. 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.
DNA ; Drug Therapy ; Glass ; Mass Screening ; Mycobacterium tuberculosis* ; Mycobacterium* ; Point Mutation* ; Rifabutin ; Rifampin ; Tuberculosis

One-third of the world population is infected with Mycobacterium tuberculosis, and tuberculosis (TB) remains a major cause of morbidity and mortality worldwide. The prevalence of TB is still increasing worldwide, and the practical guidelines for the management of TB are necessary for both national TB programs (NTP) and privatesector medical professionals. However, the recommendations on how to approach the TB control are different according to the epidemiologic and economic conditions. Health care providers should understand the differences in the approaches used and the underlying reasons so as to be better qualified to treat TB. The guideline in the United States is a representative of those in high-income and low-incidence countries, whereas the World Health Organization (WHO) guideline usually targets the populations in low-income and high-incidence countries. South Korea has two guidelines each for NTP and private-sector medical professionals. The aim of this review is to compare the guidelines for TB management and to understand the differences and the underlying reasons. The guidelines of TB management are different across Korea, United States, and WHO. There are many issues to be solved in the Korean guidelines, which need an evidence-based update based on Korean data for both NTP and private-sector medical professionals.
Diagnosis ; Health Personnel ; Humans ; Korea* ; Mortality ; Mycobacterium tuberculosis ; Prevalence ; Tuberculosis* ; United States* ; World Health Organization

Background: Rifampicin(RFP) is a key component of the antituberculous short-course chemotherapy and the RFP-resistance is a marker of multi-drug resistant(MDR) M. tuberculosis. rpoB gene encodes the beta-subunit of RNA polymerase of M. tuberculosis which is the target of RFP. Recent reports show that rpoB gene mutations are the cause of RFP resistance of M. tuberculosis and the main mechanism of rpoB gene mutation is point mutation. And PCR-SSCP is a rapid and easy method for detecting point mutations. So we performed PCR-SSCP of rpoB gene of M. tuberculosis and compared the result with traditional RFP sensitivity test. Method: The 27 RFP sensitive M. tuberculosis culture isolates and 25 RFP resistant isolates were evaluated. The RFP sensitivity test was done at the Korean Tuberculosis Istitute. The DNA was extracted by bead beater method and was amplified with primers TR-8 and TR-9 in a 20ul PCR reaction containing O.1ul(luCi) [alpha - 32p] -dCTP. After amplification, SSCP was done using non-denaturating polyacrylamide gel electrophoresis. Then direct sequencing was done in cases of different eletrophoretic mobility compared with that of H37Rv. In 19 cases, we compared PCR-SSCP results with patient's clinical course and the results of traditional RFP sensitivity test. Results: 1) All 27 RFP sensitive M. tuberculosis isolates showed the same electrophoretic mobility compared with that of H37Rv. And all 25 RFP resistant M. tuberculosis isolates showed different electrophoretic mobility. 2) The mechanism of rpoB gene mutation of M. tuberculosis is mainly point mutation. 3) The PCR-SSCP results correlate well with traditional RFP sensitivity and patient's clinical response to antituberculous treatment. Conclusion: The PCR-SSCP of rpoB gene is a very sensitive and rapid mehod in detecting RFP- resistant M. tuberculosis.
DNA ; DNA-Directed RNA Polymerases ; Drug Therapy ; Electrophoresis, Polyacrylamide Gel ; Mycobacterium ; Point Mutation ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Rifampin ; Tuberculosis

No abstract available.
Lung Neoplasms* ; Lung*

No abstract available.
Mesothelioma* ; Pleura*

Until recently, the tuberculin skin test (TST) has been the only tool available for diagnosing a latent TB infection. However, the development of new diagnostic tools, using the Mycobacterium tuberculosis (MTB)-specific early secreted antigenic target 6 (ESAT-6) and culture filtrate protein 10 (CFP-10) antigens, should improve the control of tuberculosis (TB) by allowing a more accurate identification of a latent TB infection (LTBI). Antigen-specific interferon-gamma (IFN-gamma) assays have greater specificity in BCG-vaccinated individuals, and as less biased by nontuberculous mycobacterial infections. Many comparative studies have suggested that those assays have a higher specificity than the TST, and the sensitivity of these assays are expected to remarkably improved if more MTB-specific antigens can become available. Nevertheless, the major obstacle to the widespread use of these tests is the limited financial resources. Similar to other diagnostic tests, the predictive value of IFN-gamma assays depends on the prevalence of a MTB infection in the population being tested. Therefore, prospective studies will be meeded to establish the applicability of these new assays at multiple geographic locations among patients of different ethnicities, and to determine if the IFN-gamma responses can indicate those with a high risk of progressing to active TB.
Bias (Epidemiology) ; Diagnosis* ; Diagnostic Tests, Routine ; Geographic Locations ; Humans ; Interferon-gamma* ; Latent Tuberculosis ; Mycobacterium tuberculosis* ; Mycobacterium* ; Prevalence ; Skin Tests ; Tuberculin ; Tuberculin Test ; Tuberculosis

Patients with immune-mediated inflammatory diseases (IMIDs) are increasingly being treated with anti-tumor necrosis factor (TNF) agents and are at increased risk of developing tuberculosis (TB). Therefore, diagnosis and treatment of latent TB infection (LTBI) is recommended in these patients due to the initiation of anti-TNF therapy. Traditionally, LTBI has been diagnosed on the basis of clinical factors and a tuberculin skin test. Recently, interferon-gamma releasing assays (IGRAs) that can detect TB infection have become available. Considering the high-risk of developing TB in patients on anti-TNF therapy, the use of both a tuberculin skin test and an IGRA should be considered to detect and treat LTBI in patients with IMIDs. The traditional LTBI treatment regimen consisted of isoniazid monotherapy for 9 months. However, shorter regimens such as 4 months of rifampicin or 3 months of isoniazid/rifampicin are increasingly being used to improve treatment completion rates. In this review, the screening methods for diagnosing latent and active TB before anti-TNF therapy in patients with IMIDs will be briefly described, as well as the current LTBI treatment regimens, the recommendations for managing TB that develops during anti-TNF therapy, the necessity of regular monitoring to detect new TB infection, and the re-initiation of anti-TNF therapy in patients who develop TB.
Diagnosis* ; Humans ; Interferon-gamma ; Interferon-gamma Release Tests ; Isoniazid ; Latent Tuberculosis* ; Mass Screening ; Necrosis ; Rifampin ; Skin Tests ; Tuberculin ; Tuberculin Test ; Tuberculosis ; Tumor Necrosis Factor-alpha

Despite global efforts to control tuberculosis (TB), multidrug-resistant TB (MDR-TB) is still a serious problem worldwide. The diagnosis of MDR-TB is based on mycobacterial culture followed by drug susceptibility testing, with results available in weeks to months. This requirement calls for rapid direct tests, especially genotypic tests, in which specimens are amplified directly for the detection of MDR-TB. The treatment of MDR-TB is challenging because of the high toxicity of second-line drugs and the longer treatment duration required compared to drug-susceptible TB. The selection of drugs in MDR-TB is based on the treatment history, drug susceptibility results, and TB drug resistance patterns in each region. Recent World Health Organization guidelines recommend the use of at least four second-line drugs (i.e., a newer fluoroquinolone, an injectable agent, prothionamide, and cycloserine or para-aminosalicylic acid) in addition to pyrazinamide. Kanamycin is the initial choice of an injectable drug, and newer fluoroquinolones include levofloxacin and moxifloxacin. For extensively drug-resistant TB, group 5 drugs such as linezolid and clofazimine need to be included. New drugs such as delamanid and bedaquiline have recently been approved for treating MDR-TB and other agents with novel mechanisms of action that can be given for shorter durations (6-12 months) for MDR-TB are under investigation.
Clofazimine ; Cycloserine ; Diagnosis* ; Drug Resistance ; Fluoroquinolones ; Kanamycin ; Levofloxacin ; Prothionamide ; Pyrazinamide ; Tuberculosis ; Tuberculosis, Multidrug-Resistant* ; World Health Organization

BACKGROUND/AIMS: The usefulness of interferon-γ release assays (IGRAs) in monitoring to responses to anti-tuberculosis (TB) treatment is controversial. We compared the results of two IGRAs before and after anti-TB treatment in same patients with active TB. METHODS: From a retrospective review, we selected patients with active TB who underwent repeated QuantiFERON-TB Gold (QFN-Gold, Cellestis Limited) and T-SPOT.TB (Oxford Immunotec) assays before and after anti-TB treatment with first-line drugs. Both tests were performed prior to the start of anti-TB treatment or within 1 week after the start of anti-TB treatment and after completion of treatment. RESULTS: A total of 33 active TB patients were included in the study. On the QFN-Gold test, at baseline, 23 cases (70%) were early secreted antigenic target 6-kDa protein 6 (ESAT-6) or culture filtrate protein 10 (CFP-10) positive. On the T-SPOT. TB test, at baseline, 31 cases (94%) were ESAT-6 or CFP-10 positive. Most of patients remained both test-positive after anti-TB treatment. Although changes in interferon-γ release responses over time were highly variable in both tests, there was a mean decline of 27 and 24 spot-forming counts for ESAT-6 and CFP-10, respectively on the T-SPOT.TB test (p < 0.05 for all). CONCLUSIONS: Although limited by the small number of patients and a short-term follow-up, there was significant decline in the quantitative result of the T-SPOT. TB test with treatment. However, both commercial IGRAs may not provide evidence regarding the cure of disease in Korea, a country where the prevalence of TB is within the intermediate range.
Follow-Up Studies ; Humans ; Interferon-gamma Release Tests ; Korea ; Prevalence ; Retrospective Studies ; Tuberculosis

The Mainz classification for renal cell tumors was introduced in 1986 and it's utility has been reported in several histogenetic and genetic studies of renal cell tumors. We present a study of 127 cases of renal cell tumors with clinicopathologic correlation, DNA content analysis, and histogenesis studied by histochemical and immunohistochemical staining. The 127 renal cell tumors classified by the Mainz classification were 87 clear cell, 17 chromophilic, 13 chromophobe and 3 sarcomatoid renal cell carcinomas, 5 oncocytomas and 2 adenomas. These subtypes showed significant correlation not with age, sex, Robson's stage, DNA ploidy or tumor recurrence but with nuclear grade (p=0.001) and tumor size (p=0.001). Hall's colloidal iron (p=0.002) and carbonic anhydrase II (p=0.013) stains, representing the origin of distal nephron especially of collecting duct, were significantly correlated with specific subtypes of renal cell tumors, especially chromophobe cell renal carcinoma. This study demonstrates that the Mainz classification suggests several morphologically different subtypes and variants of renal cell tumors and that some of them may have originated from the distal nephron, particularly from the collecting duct.
Adenoma ; Adenoma, Oxyphilic ; Carbonic Anhydrase II ; Carcinoma, Renal Cell ; Classification* ; Colloids ; Coloring Agents ; DNA* ; Iron ; Nephrons ; Ploidies ; Recurrence