ISID's research grants are a key part of the Society’s efforts to combat infectious diseases, especially in countries facing infectious diseases burden the most and economic challenges. We are excited to share the progress of three mentees from Cohort II as they advance their research projects!

Hombamane Christelle Simfele, Togo

Project: Molecular Characterisation of Single-Nucleotide Polymorphisms (SNPs) associated with Resistance of Ancylostoma Duodenale to Albendazol in Hookworm-Infected Individuals in Togo

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We analysed the data related to the determination of single-nucleotide polymorphisms associated with resistance of Ancylostoma duodenale to albendazole in infected subjects using RT-PCR test. For that:

To determine whether real-time PCR was able to detect mutations in codon positions 167 and 200 of beta-tubulin isotype-1, plasmids (control clones) for the resistance alleles were designed using a site-directed mutagenesis approach. These ‘resistant clone’ plasmids were then tested at different concentrations by PCR with primers specific (resistant or susceptible) to A. duodenale. PCR reactions containing resistance controls and using primers specific for the resistant SNP amplified products with an earlier threshold than PCR reactions using the same DNA templates but containing primers specific for the susceptible allele. The 1000ng/uL concentration of the resistant plasmid showed an optimal Ct value for detection (Ct= 12.29) compared to the other concentrations.

After assessing the efficacy of the controls and determining the optimum concentration to use, real-time PCR tests were then performed to examine DNA samples from subjects infected with A. duodenale. A total of 16 DNA samples were examined. For all samples tested, the average percentage of susceptible alleles at codon 167 of the beta-tubulin isotype-1 gene was 76.51% and the percentage of resistant alleles was 23.48%. At codon 200, the frequency of sensitive alleles was 99.67% and that of resistant alleles was 0.32%. These results show the presence of resistance markers, particularly at codon 167 of the beta-tubulin gene in hookworm-infected subjects.

Based on calculations of the proportion of single nucleotide polymorphisms (SNPs) for codon 167 and codon 200 in subjects infected with A. duodenale, the frequency of SNPs for codon 167 was significantly higher than that for codon 200 (p<0.0001), The quality control of reagents delivered has been performed using specific tests such as PCR tests for Primers and plasmids (Positive controls)

We have started our second objective which was to sequence samples for the detection of SNPs in Ancylostoma duodenale positive samples at the University of Ghana, to do this, authorisation was obtained for integration into the molecular biology laboratory at the University of Ghana (WACCBIP). We went to the university with the samples for a period of 1 month to carry out the sequencing on the samples.

The first phase of this sequencing stage involved assessing the efficiency of the samples by measuring the concentration of DNA contained in the samples using the nanodrop and Qubit. The conventional PCR was then optimised to obtain the correct size bands corresponding to target the beta-tubulin isotype-1 gene.

Measurement of sample concentration revealed that all samples had sufficient concentrations to perform targeted sequencing for the β-tubulin gene. Conventional PCR revealed unexpected non-specific bands. However, optimisations are underway to obtain bands specific to the β-tubulin gene.

Matilda Kamara, Sierra Leone

Project: Antibiotic use and resistance in a slum community in Freetown: an analysis of gender intersectionality in a human-environment interface

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The aims of our projects are:

Aim 1: To assess antibiotic self-prescription and community perception about AMR. We disaggregate data to understand gender intersectionality with other demographic and socio-economic dimensions in antibiotic self-prescription and community perception about AMR. We hypothesize that the intersection of gender with other dimensions has a significant influence on antibiotic self-prescription and community perception about AMR, but this is not currently known. We will know more about this intersection with our study. The proposed output for this aim is a manuscript defining gender intersectionality with other dimensions on antibiotic self-prescription and community perception about AMR in a slum community in Western Sierra Leone.

Aim 2: To assess the quality of drinking water sources of this slum community by assessing the levels of E. coli and related antibiotic resistance. We hypothesize that the sources of drinking water in this slum community have a high load of resistance E. coli. The proposed output is a manuscript on resistance E. coli in drinking water containing actionable recommendations on

We are advancing to accomplish most of our project goals. For the qualitative arm of the study we have completed the transcription and translation of key informant interviews and for the quantitative arm we have completed data analysis. We have also been able to collect water samples and analyze them through culture and sensitivity testing.

Mawuli Gohoho, Ghana

Project: Optimizing TB Case Notification through Capacity Building: An Assessment of the Accuracy and Efficiency of the Revised TB Screening Tool Roll-out in Jasikan Municipality, Ghana

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After ERB approval, baseline data collection across the facilities was started and completed. The mHealth App for presumed TB referral has also been finalized. The piloting showed a few challenges which were also addressed. Stakeholder engagement and community entry were expected to be done, but due to other conflicting public health programs and reactive campaigns implementation in the nation which involved the district, utilizing same resources as the project, the activity was not fruitful.

For the next period, Stakeholder engagement on the intervention and capacity building on the App usage are planned to be conducted, representing approximately 60% achievement in this reporting period.

Fig 1: These photos above capture TB Screening Supervisors interacting with trained healthcare workers (both preventive and curative) in the Jasikan Municipality, assessing their screening registers and providing necessary on-the-job coaching as well.