Development, assessment, an implementation of a dual isothermal point-of care molecular diagnostic for both FGS and HPV

FGS is a gynaecological condition resulting from infection with the parasite Schistosoma haematobium and is thought to affect over 40 million women living across sub-Saharan Africa. In addition to causing symptoms such as bloody urine, severe cramping, and infertility, S. haematobium infection is associated with heightened risk of STI's and is a Grade 1 carcinogen.

HPV is group of viruses known to cause genital warts and is often a precursor to cervical cancer. HPV is a global pathogen, and high-risk HPV strains cause more than 90% of all cervical cancer cases. A major hurdle in successful diagnosis and treatment in for Least Developed Countries (LDC's) is the lack of accessible diagnostics.

Due to the fact that both HPV and FGS can be detected through gynaecological samples, this project aims to provide an integrated diagnostic that can be implemented in low resource settings to support timely treatment of marginalised women.

Project Plan
1. To review current literature to identify isothermal molecular assays that have been developed and used for the diagnosis of FGS and HPV. These assays will also be reviewed in terms of their applicability in low resource settings.

2. Suitable targets and primer-probe sets for RPA and LAMP assay development will be selected or redesigned (if needed) using genetic data from open-access databases. Online resources, including BLAST and OligoAnalyzer, will be used to in silico test the specificity of the assays followed by laboratory analytical development and testing. For HPV, a general HPV target and a high-risk HPV target would both be investigated.

3. Assays will be tested and optimised as singleplex reactions before being optimised in a multiplex.

4. An internal control target (human DNA) will be investigated and aimed to be incorporated into the assay for quality control purposes.

5. Analytical testing will be done using synthetic DNA, gDNA from reference samples, and also available clinical samples for both S. haematobium and HPV. All molecular tests will be validated against qPCR reference tests.

6. The assays we be evaluated in terms of their fit to the REASSURED criteria set out for diagnostics from the WHO for diagnostics for LDC's.

7. Final stage assessment of the assay performance and suitability will be performed in a S. haematobium / HPV co-endemic setting in Sub-Saharan Africa. A comparison between self-swab and clinical swab samples will also be made to help inform the acceptability of the sampling procedure within the target population. Considering cultural and social attitudes is essential in relation to the most accurate diagnosis

8. As well as descriptive statistics, differences in diagnostic capabilities between qPCR and the developed multiplex assay would be carried out using latent class analysis.

The project will develop interdisciplinary skills including molecular biology, chemistry, field work, epidemiological analysis, and statistical analysis. Knowledge of physiology and pathology, skills identified as vulnerable by the UKRI, will also be developed.

There is emphasis on intercollegiate collaboration, with significant portions of the project being undertaken at each institution due the bespoke expertise of each partner. Additionally, existing connections with commercial diagnostic companies and non-for-profit organisations that are dedicated to supporting work towards the provision of accessible diagnostics to marginalized communities, will guide this research.

The project considers the MRC LID themes of global health and infectious disease, which are essential to the MRC strategy of improving human health and economic prosperity for everyone. Schistosomiasis is a neglected tropical disease and is most prevalent in LDC's. HPV is present globally, but access to screening and vaccination is limited in LDCs.