Browsing by Author "Wasswa, Ronald"

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    Comparison of non-magnetic and magnetic beads multiplex assay for assessment of Plasmodium falciparum antibodies
    (PeerJ, 2019-01-03) Ondigo, Bartholomew N.; Park, Gregory S.; Ayieko, Cyrus; Nyangahu, Donald D.; Wasswa, Ronald; John, Chandy C.; Biochemistry and Molecular Biology, School of Medicine
    Background: New reagents have emerged allowing researchers to assess a growing number of vaccine-associated immune parameters. Multiplex immunoassay(s) are emerging as efficient high-throughput assays in malaria serology. Currently, commercial vendors market several bead reagents for cytometric bead assays (CBA) but relative performances are not well published. We have compared two types of bead-based multiplex assays to measure relative antibody levels to malarial antigens. Methods: Assays for the measurement of antibodies to five Plasmodium falciparum vaccine candidates using non-magnetic and magnetic fluorescent microspheres were compared for their performances with a Bio-Plex200 instrument. Mean fluorescence intensity (MFI) was determined from individuals from western Kenya and compared to known positive and negative control plasma samples. Results: P. falciparum recombinant antigens were successfully coupled to both non-magnetic and magnetic beads in multiplex assays. MFIs between the two bead types were comparable for all antigens tested. Bead recovery was superior with magnetic beads for all antigens. MFI values of stored non-magnetic coupled beads did not differ from freshly coupled beads, though they showed higher levels of bead aggregation. Discussion: Magnetic and non-magnetic beads performed similarly in P. falciparum antibody assays. Magnetic beads were more expensive, but had higher bead recovery, were more convenient to use, and provided rapid and easy protocol manipulation. Magnetic beads are a suitable alternative to non-magnetic beads in malarial antibody serology.
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    Severe Anemia Is Associated with Systemic Inflammation in Young Children Presenting to a Tertiary Hospital in Uganda
    (American Society of Tropical Medicine and Hygiene, 2020-12) Opoka, Robert O.; Conroy, Andrea L.; Waiswa, Ali; Wasswa, Ronald; Tumwine, James K.; Karamagi, Charles; John, Chandy C.; Pediatrics, School of Medicine
    The role of inflammation in severe anemia (SA) in African children has not been well characterized. We conducted a study to evaluate risk factors for SA in young children admitted at a tertiary unit in Uganda. Clinical, infectious, and micronutrient risk factors for anemia, along with markers of inflammation, were evaluated in children aged < 5 years in Jinja Hospital, Uganda. Participants included 284 children with SA (Hemoglobin [Hb] < 5.0 g/dL), and two control groups: 63 children admitted with acute illness without SA (Hb > 9.3 g/dL) and 53 asymptomatic community control children. Appropriate logistic analysis was performed to determine factors associated with SA. Of the 284 children with SA, 36.5% had Plasmodium falciparum parasitemia, 32.7% had blackwater fever (one of the types of severe malaria), and 15.5% had vitamin B12 deficiency. HIV infection, bacteremia, hookworm infection, severe acute malnutrition, and folate deficiency were relatively uncommon (each accounting for < 8%). Factors independently associated with SA compared with the combined control groups included (adjusted odds ratio [OR]; 95% CI) the following: P. falciparum parasitemia (OR: 4.3; 95% CI: 1.4-13.8), total white blood count (OR: 1.3; 95% CI: 1.1-1.4), C-reactive protein (OR: 1.8; 95% CI: 1.3-2.4), and ferritin (OR: 2.7; 95% CI: 1.9-4.0). In this area of Uganda, malaria and markers of inflammation were independently associated with SA in children. Additional studies are required to determine the role of inflammation in children with SA in this population.