Protocols

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Browsing Protocols by Subject "Research Subject Categories::MEDICINE"

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    Hybridization in urogenital schistosomiasis (HUGS)
    (Kamuzu University of Health Sciences, 2021-10-18) Musaya, Janelisa
    Type of study: Cohort study, with cross-sectional snail surveys Problem to be studied: Schistosomiasis is a Neglected Tropical Disease (NTD) caused by infection with parasitic flatworms of the Schistosoma genus. With our discovery of novel Schistosoma haematobium hybrids commonly co-infecting Malawian school children, critical knowledge gaps in the biology and control of schistosomiasis in Central Africa have emerged. These amplify formal concerns of the WHO’s preventive chemotherapy (PC) strategy and underpinning targets within their 2021-2030 Roadmap. Our HUGS (hybridisation in urogenital schistosomiasis) investigation aims to address these critical knowledge gaps in the transmission biology, applied epidemiology and clinical importance of S. haematobium hybrids. Objectives: The objectives of the study are: i. To test if the proportion of hybrid coinfection is uniform across two representative communities in Mangochi and Nsanje districts where S. haematobium-mattheei or S. haematobiumbovis occur, inclusive of household GPS mapping and identification of associated risk factors by questionnaire. ii. To verifyassess any changes in the proportions and spatial patterns of hybrid coinfection with annual praziquantel treatment, in a 2-year longitudinal population follow-up studystudy, if the above proportions and spatial patterns of hybrid coinfection hold, or alter, with annual praziquantel treatment. iii. To ascertain if there is any altered host morbidity (e.g. urogenital inflammation) in hybrid coinfection(s) as measured by portable ultrasonography and point-of-contact assays. iv. To reveal describe the hybrid environmental transmission dynamics of the hybrid coinfection(s) upon malacological, livestock tracking and abattoir surveys, dissecting schistosome material by advanced genetic profiling. Methodology: This multi-study will comprise of environmental snail surveys, animal-abattoir surveys and human cohort studies. Twelve quarterly spaced environmental snail surveys will commence before, during and complete after, examinations for hybrids in livestock and people to give best chance of revealing environmental transmission. Three inspections of slaughtered livestock at abattoir, each precede human sampling to better alert novel hybrids/animal schistosomes before three annual examinations of the human cohort, baseline and two follow-up surveys. Data will be obtained from a questionnaire and participants will also provide urine and stool samples on the same day which will be tested for Schistosoma-hybrids. Expected findings and dissemination: We will describe the prevalence of Schistosomahybrids. We will therefore share our findings with the Kamuzu University of Health Sciences, (COMREC), Mangochi District Health Office,District Health Management Team, Communities in Mangochi, the District Commissioner and District Executive Committee. We will also present findings at local and international conferences and in peer reviewed publications.
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    Investigating effects of topoisomerase mutations on topoisomeraseciprofloxacin interaction in Salmonella typhi isolated in Blantyre
    (Kamuzu Universiyt of Health Sciences, 2021-10-06) Matambo, Ernest
    Introduction Typhoid fever accounts for about 135, 000 deaths in adults and children in south Asia, central and southern Africa. The recommended antibiotic treatment of typhoid fever in Malawi is the fluoroquinolone ciprofloxacin, however, Salmonella enterica subspecies enterica serovar Typhi (S. Typhi) decreased susceptibility to ciprofloxacin has been detected in Blantyre, Malawi. S. Typhi can become resistant to fluoroquinolones by two mechanisms; either due to the acquisition of plasmid mediated resistance genes or mutations in gyrA, gyrB, parC and parE genes in topoisomerase II and IV. This study will use in silico methods to identify mutations in gyrA, gyrB, parC and parE genes in S. Typhi from Blantyre and to investigate the effects of topoisomerase II and IV mutations on topoisomerase-ciprofloxacin interaction. Main objective To investigate the effect of gyrA, gyrB, parC and parE mutations on ciprofloxacintopoisomerase interaction in S. Typhi isolated in Blantyre. Specific objectives To identify mutations in Quinolone Resistance Determining Region of S. Typhi with decreased ciprofloxacin susceptibility isolated in Blantyre. To describe effects of mutations in gyrA, gyrB, parC and parE genes on their protein structure in S. Typhi with decreased ciprofloxacin susceptibility isolated in Blantyre. To compare folding energy of topoisomerase protein structure of S. Typhi with decreased ciprofloxacin susceptibility to topoisomerase protein structure of ciprofloxacin-sensitive S. Typhi. To compare ciprofloxacin-topoisomerase interaction in topoisomerase of S. Typhi with decreased ciprofloxacin susceptibility to ciprofloxacin-sensitive S. Typhi Methodology This will be a retrospective study. Reads of S. Typhi with reduced susceptibility to ciprofloxacin (n=14) and of ciprofloxacin sensitive S. Typhi (n=14) will be assembled using de novo assembly. Topoisomerase II and IV genes will then be extracted from the assembled genomes. Topoisomerase II and IV gene mutations will be identified in the genomes of S. Typhi with decreased susceptibility to ciprofloxacin by comparing the 06-Oct-2021 Ver1.1 September 2021 gene sequences to the CT18 reference genome. The topoisomerase II and IV gene sequences will be translated into protein sequences and then their protein structures predicted. The protein structures will be analysed for structural differences in the S. Typhi with decreased ciprofloxacin susceptibility and ciprofloxacin-sensitive S. Typhi. Folding energy between S. Typhi with decreased ciprofloxacin susceptibility and ciprofloxacin-sensitive S. Typhi protein structures will be compared. Bonding energy with ciprofloxacin between the two groups of protein structures will also be compared. Expected findings and their dissemination Expected findings are that there will be mutations in topoisomerase II and IV gene sequences and that the mutations will be in the ciprofloxacin binding pockets of the predicted protein structures in the S. Typhi with decreased ciprofloxacin susceptibility. Folding energy is expected to be low in protein structures without mutations as compared to protein structures with mutations. Protein structures with mutations will have high binding energy as compared to those without mutations. The study findings and write up will be submitted to COMREC, Kamuzu University of Health Sciences Library and to a peer-reviewed journal for publication. 06