Local Manufacturing of Enzymes
For The Multiplex Diagnosis of
Candidiasis and STIs
The burden of Candidiasis and Sexually Transmitted Infections (STIs) still remains a major health challenge in Ghana, a Low- and Middle-Income Country (LMIC) in Sub-Saharan Africa. For example, candidiasis caused by Candida albicans currently accounts for over 70% of neonatal sepsis and death in Ghana.
The diagnosis of Candidiasis, Neisseria gonorrhoea, Syphilis, and Trichomoniasis in Ghana still relies on microscopy and serology respectively. However, these methods lack sensitivity and specificity, take several days to weeks to complete and are error prone. Molecular-based diagnostics detecting DNA are sensitive and specific and take only a few hours to complete. However, they are not currently used in mainstream diagnosis and one reason is the need for reagents such as enzymes that can be costly and challenging to ship.
This project seeks to locally biomanufacture two high fidelity DNA polymerases: KOD and Pfu-Sso7d (commonly known as Phusion). They will be expressed in a bacterial host, purified using novel self-aggregating tags removing the need for complex downstream processing and incorporated into a tailored protocol for multiplex PCR-molecular diagnosis of Candida albicans, Neisseria gonorrhoeae, Treponema pallidum and Trichomonas vaginalis for use in Ghanaian clinics and hospitals.
It is expected that at the end of this study, we will have demonstrated the local production of enzymes and their use in a proof-of-concept novel multiplex PCR format that is validated with positive control samples and clinical samples. This will be the starting point in developing affordable test kits for diagnosis of candidiasis and STIs.