Browsing by Author "Lubega, G.W."

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    In Vitro Trypanocidal Activity of Antibodies to Bacterially Expressed Trypanosoma brucei Tubulin
    (Iranian Journal of Parasitology, 2012) Kateete, D.P.; Alezuyo, C.; Nanteza, A.; Lubega, G.W.
    There are only four drugs for treating African trypanosomiasis, a devastating disease in sub-Saharan Africa. With slow discovery of better drugs, vaccination is viewed as the best method of control. We previously showed that antibodies to native Trypanosoma brucei brucei tubulin inhibit the growth of trypanosomes in culture. Here, we aimed to determine the effect of antibodies to bacte­rially expressed trypanosome tubulin on T. brucei brucei growth. T. brucei brucei alpha and beta tubulin genes were individually expressed in Escherichia coli under the tryptophan promoter. Monoclonal tubulin antibodies reacted specifically with the ex­pressed tubulins with no cross-reaction with the opposite tubulin. Rabbits were immunized with 450μg each of the concentrated recombinant tubulin, and production of antibodies assessed by ELISA and Western blotting. The effect of polyclonal antibodies on trypanosome growth was deter­mined by culturing bloodstream T. brucei brucei in up to 25% of antisera. Low antisera dilutions (25%) from the immunized rabbits inhibited trypanosome growth. The most cytotoxic antisera were from one rabbit immunized with a mixture of both alpha and beta tubulins. However, the result was not reproduced in other rabbits and there was no apparent effect on growth at higher antisera dilutions. Antibodies to bacterially expressed trypanosome tubulin are not effective at killing cul­tured bloodstream trypanosomes.
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    Using Translation Elongation Factor Gene to Specifically Detect and Diagnose Fusarium xylaroides, a Causative Agent of Coffee Wilt Disease in Ethiopia, East and Central Africa
    (J Plant Pathol Microbiol, 2018) Olal, S.; Olango, N.; Kiggundu, A.; Ochwo, S.; Adriko, J.; Nanteza, A.; Matovu, E.; Lubega, G.W.; Kagezi, G.; Hakiza, G.J.; Wagoire, W.W.; Opiyo, S.O.
    The present study presents the first report on the application of DNA-based polymerase chain reaction (PCR) for the specific detection and diagnosis of F usarium xylarioides (anamorph: G ibberrela xylarioides). Fusarium xylarioides is the causative agent of Coffee wilt disease (Tracheomycosis), and the disease is the most important economic constraint in Robusta coffee production in Uganda. The pathogen has two races, one pathogenic to Robusta coffee and the other to Arabica coffee, and not vice versa. Its laboratory diagnosis has been mainly based on microscopy, which is slow, has poor discriminative power, requires high expertise, only applicable on host plants with symptoms, and has since failed to detect the pathogen from the soil. Translation Elongation factor-1α (TEF-1α) gene from a F. xylarioides isolated from infected Robusta coffee plant was amplified by Fusarium genus specific primer then the PCR product sequenced. The sequence data was then used to design the specific primer. The primer-BLAST product was found to match only F. xylarioides sequences comprising 75% of the race pathogenic to Robusta and 25% to Arabica coffee. In vitro test by PCR showed the primer to be specific to only F. xylarioides amplifying a 284bp product and was able to differentiate F. xylarioides from all closely related species of Fusarium and other plant pathogens tested. More so it was able to amplify DNA from all the F. xylarioides isolates from different regions of Uganda, and amplified DNA concentrations as minute as 0.78 ng/µL.