Browsing by Author "Ogwang, Sam"

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    Anti-tubercular activity of Callistemon citrinus and Piptadenistrum africanum on resistant strains of Mycobacterium tuberculosis using Microplate alamar blue assay
    (ScopeMed, 2015) Bunalema, Lydia; Tabuti, John; Sekagya, Yahaya; Ogwang, Sam; Waako, Paul
    Tuberculosis (TB) is one of the leading causes of death among infectious diseases in the world. It is responsible for killing approximately 1.4 million people per year worldwide. This devastating situation has steadily worsened, exacerbated by the emergence of drug-resistance and Human Immunodeficiency Virus (HIV) co-infection. The objectives of the study were to determine the minimum inhibitory concentration (MIC) of selected plant species on three TB strains and to determine different phytochemicals contained in the plant species. Methods: Microplate Alamar Blue Assay (MABA) was used to determine the MIC of two commonly mentioned plant species, Piptadenistrum africanum and Callistemon citrinus on resistant variant strains of Mycobacterium tuberculosis. Qualitative tests were used to determine the phytochemicals in the plants. Results: The chloroform extract of Callistemon spp. had MICs of 0.048mg/ml, 0.158mg/ml and 0.19mg/ml on the pan sensitive, isoniazid resisistant and rifampicin resistant strains respectively. P. africanum had MICs of 0.395 mg/ml, 0.395 mg/ml and 0.78 mg/ml on the pan sensitive, rifampicin and isoniazid resistant strains respectively. Conclusion: These plant species appear to be active not only on the pan sensitive strains of TB but also on resistant strains and could be developed into drugs for the treatment of Multi drug resistant (MDR) TB.
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    Comparison of MGIT and Myco/F Lytic Liquid-Based Blood Culture Systems for Recovery of Mycobacterium tuberculosis from Pleural Fluid
    (Journal of clinical microbiology, 2015) Harausz, Elizabeth; Kafuluma Lusiba, John; Nsereko, Mary; Johnson, John L.; Toossi, Zahra; Ogwang, Sam; Boom, Henry; Joloba, Moses L.
    Tuberculosis (TB) is the most frequent cause of exudative pleural effusions in areas of high TB incidence. Studies have shown that Mycobacterium tuberculosis is the causative agent in up to 44% of HIV-seronegative people hospitalized with a pleural effusion (1–3), and the percentage is higher in HIV-seropositive people (4). Pleural TB is a paucibacillary disease. The pathogenesis of a tuberculous pleural effusion is likely due to a delayed hypersensitivity reaction to M. tuberculosis proteins (for a review, see reference 5) and not to a large burden of organisms. The scarcity of organisms makes it difficult to isolate M. tuberculosis from pleural fluid samples, leading to low rates of culture confirmation. Rich culture media are generally more sensitive in detecting M. tuberculosis in sputum and other clinical samples (6). Few studies have compared different liquid media and examined their potential role in combination with solid media for the diagnosis of tuberculous pleurisy. In this study, we compared the Bactec 9120 Myco/F lytic blood culture system (Myco/F lytic) to the Bactec mycobacterial growth indicator tube (MGIT) 960 system (Becton Dickinson, Sparks, MD) (with each liquid system used in conjunction with locally prepared Middlebrook 7H11 solid medium) with respect to time to positivity (TTP), sensitivity, specificity, and percent culture yield of M. tuberculosis isolates from pleural fluid.
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    Comparison of rapid tests for detection of rifampicin-resistant Mycobacterium tuberculosis in Kampala, Uganda
    (BMC infectious diseases, 2009-08-26) Ogwang, Sam; Asiimwe, Benon B.; Okwera, Alphonse; Joloba, Moses L.
    Drug resistant tuberculosis (TB) is a growing concern worldwide. Rapid detection of resistance expedites appropriate intervention to control the disease. Several technologies have recently been reported to detect rifampicin resistant Mycobacterium tuberculosis directly in sputum samples. These include phenotypic culture based methods, tests for gene mutations and tests based on bacteriophage replication. The aim of the present study was to assess the feasibility of implementing technology for rapid detection of rifampicin resistance in a high disease burden setting in Africa. Sputum specimens from re-treatment TB patients presenting to the Mulago Hospital National TB Treatment Centre in Kampala, Uganda, were examined by conventional methods and simultaneously used in one of the four direct susceptibility tests, namely direct BACTEC 460, Etest, "in-house" phage test, and INNO- Rif.TB. The reference method was the BACTEC 460 indirect culture drug susceptibility testing. Test performance, cost and turn around times were assessed. In comparison with indirect BACTEC 460, the respective sensitivities and specificities for detecting rifampicin resistance were 100% and 100% for direct BACTEC and the Etest, 94% and 95% for the phage test, and 87% and 87% for the Inno-LiPA assay. Turn around times ranged from an average of 3 days for the INNO-LiPA and phage tests, 8 days for the direct BACTEC 460 and 20 days for the Etest. All methods were faster than the indirect BACTEC 460 which had a mean turn around time of 24 days. The cost per test, including labour ranged from $18.60 to $41.92 (USD). All four rapid technologies were shown capable of detecting rifampicin resistance directly from sputum. The LiPA proved rapid, but was the most expensive. It was noted, however, that the LiPA test allows sterilization of samples prior to testing thereby reducing the risk of accidental laboratory transmission. In contrast the Etest was low cost, but slow and would be of limited assistance when treating patients. The phage test was the least reproducible test studied with failure rate of 27%. The test preferred by the laboratory personnel, direct BACTEC 460, requires further study to determine its accuracy in real-time treatment decisions in Uganda.
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    Rate and Amplification of Drug Resistance among Previously-Treated Patients with Tuberculosis in Kampala, Uganda
    (Clinical infectious diseases, 2008) Temple, Beth; Ayakaka, Irene; Ogwang, Sam; Nabanjja, Helen; Kayes, Susan; Nakubulwa, Susan; Worodria, William; Levin, Jonathan; Joloba, Moses; Okwera, Alphonse; Eisenach, Kathleen D.; McNerney, Ruth; Elliott, Alison M.; Smith, Peter G.; Mugerwa, Roy D.; Ellner, Jerrold J.; Jones-Lopez, Edward C.
    Drug-resistant Mycobacterium tuberculosis has emerged as a global threat. In resource-constrained settings, patients with a history of tuberculosis (TB) treatment may have drug-resistant disease and may experience poor outcomes. There is a need to measure the extent of and risk factors for drug resistance in such patients. Methods. From July 2003 through November 2006, we enrolled 410 previously treated patients with TB in Kampala, Uganda. We measured the prevalence of resistance to first- and second-line drugs and analyzed risk factors associated with baseline and acquired drug resistance. Results. The prevalence of multidrug-resistant TB was 12.7% (95% confidence interval [95% CI], 9.6%–16.3%). Resistance to second-line drugs was low. Factors associated with multidrug-resistant TB at enrollment included a history of treatment failure (odds ratio, 23.6; 95% CI, 7.7–72.4), multiple previous TB episodes (odds ratio, 15.6; 95% CI, 5.0–49.1), and cavities present on chest radiograph (odds ratio, 5.9; 95% CI, 1.2–29.5). Among a cohort of 250 patients, 5.2% (95% CI, 2.8%–8.7%) were infected with M. tuberculosis that developed additional drug resistance. Amplification of drug resistance was associated with existing drug resistance at baseline (P ! .01) and delayed sputum culture conversion (P ! .01). Conclusions. The burden of drug resistance in previously treated patients with TB in Uganda is sizeable, and the risk of generating additional drug resistance is significant. There is an urgent need to improve the treatment for such patients in low-income countries.
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    Retrospective Analysis of Archived Pyrazinamide Resistant Mycobacterium tuberculosis Complex Isolates from Uganda—Evidence of Interspecies Transmission
    (Microorganisms, 2019) Wanzala, Sylvia I.; Nakavuma, Jesca; Travis, Dominic; Kia, Praiscillia; Ogwang, Sam; Waters, Wade R.; Thacker, Tyler; Johnson, Timothy; Hadi, Syeda A.; Sreevatsan, Srinand
    The contribution of Mycobacterium bovis to the proportion of tuberculosis cases in humans is unknown. A retrospective study was undertaken on archived Mycobacterium tuberculosis complex (MTBC) isolates from a reference laboratory in Uganda to identify the prevalence of human M. bovis infection. A total of 5676 isolates maintained in this repository were queried and 136 isolates were identified as pyrazinamide resistant, a hallmark phenotype of M. bovis. Of these, 1.5% (n = 2) isolates were confirmed as M. bovis by using regions of di erence PCR analysis. The overall size of whole genome sequences (WGSs) of these two M. bovis isolates were ~4.272 Mb (M. bovis Bz_31150 isolated from a captive chimpanzee) and 4.17 Mb (M. bovis B2_7505 from a human patient), respectively. Alignment of these genomes against 15 MTBC genome sequences revealed 7248 single nucleotide polumorphisms (SNPs). Theses SNPs were used for phylogenetic analysis that indicated a strong relationship between M. bovis and the chimpanzee isolate (Bz_31150) while the other M. bovis genome from the human patient (B2_7505) analyzed did not cluster with any M. bovis or M. tuberculosis strains. WGS analysis also revealed multidrug resistance genotypes; these genomes revealed pncA mutations at positions H57D in Bz_31150 and B2_7505. Phenotypically, B2_7505 was an extensively drug-resistant strain and this was confirmed by the presence of mutations in the major resistance-associated proteins for all anti-tuberculosis (TB) drugs, including isoniazid (KatG (S315T) and InhA (S94A)), fluoroquinolones (S95T), streptomycin (rrs (R309C)), and rifampin (D435Y, a rare but disputed mutation in rpoB). The presence of these mutations exclusively in the human M. bovis isolate suggested that these occurred after transmission from cattle. Genome analysis in this study identified M. bovis in humans and great apes, suggesting possible transmission from domesticated ruminants in the area due to a dynamic and changing interface, which has created opportunity for exposure and transmission.
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    Sulfamethoxazole Susceptibility of Mycobacterium tuberculosis Isolates from HIV-Infected Ugandan Adults with Tuberculosis Taking Trimethoprim-Sulfamethoxazole Prophylaxis
    (Antimicrob Agents Chemother, 2015) Ogwang, Sam; Good, Caryn E.; Okware, Brenda; Nsereko, Mary; Jacobs, Michael R.; Boom, W. Henry; Bark, Charles M.
    Alternative drugs are urgently needed to treat multidrug-resistant (MDR) tuberculosis (TB). Given the difficulties of new drug development, repurposing currently licensed antibiotics is practical and efficient. Trimethoprim-sulfamethoxazole (SXT) is a fixed-dose drug combination used worldwide as treatment and prophylaxis for multiple infections. Sulfamethoxazole (SMX) is in the sulfonamide class of antibiotics, which were explored as an anti-TB treatment in the mid-20th century with early studies showing potential value for the treatment of pulmonary and miliary TB (1–5). More recently, Forgacs et al. reported defervescence of a patient with pulmonary TB who was initially treated with SXT alone and also demonstrated in vitro susceptibility to SXT in 43 of 44 Mycobacterium tuberculosis isolates (6). These drug susceptibility results were independently confirmed in laboratory strains (7, 8) and in patient isolates demonstrating SMX to be the active agent with MICs within achievable serum levels (9, 10). In addition, Alsaad and colleagues reported the use of SXT as part of a combination regimen used to treat 10 patients with MDR-TB in the Netherlands (11). They also reported M. tuberculosis susceptibility to SXT in 17 of 18 patients with TB-HIV coinfection; however, only 1 was taking SXT prior to TB diagnosis (12). Given the development of drug resistance when active TB is treated with a single drug, there is concern for resistance to SMX among TBHIV- coinfected patients taking SXT prophylaxis. To address this concern, we performed drug susceptibility testing (DST) on M. tuberculosis isolates obtained from pretreatment sputum specimens of HIV-infected patients taking SXT prophylaxis at the time of diagnosis of active TB. Sputum isolates used for