A rapid, accurate, and easy-to-use diagnostic assay for STIs

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PROJECT SUMMARY The aim of this Phase II proposal is to develop a rapid, easy-to-use, and inexpensive multi-sample diagnostic system to identify sexually transmitted infection (STI) pathogens, such as Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) in non-traditional healthcare settings. The features of our system will likely qualify it to be a CLIA-waived CT/NG test. The sensitivity and specificity to be achieved using this prototype type assay will be comparable with those of current laboratory-based molecular tests. This would present a major advancement in clinical diagnostics for STIs. According to the CDC, chlamydia and gonorrhea, caused by C. trachomatis and N. gonorrhoeae, are the two most frequently reported STIs in the US and likely the rest of the world. These STIs are often under-diagnosed, leading to delayed treatment, continued spread and higher public healthcare costs (~$16 billion annually). As a result of their prevalence and long term health consequences, there is a need for rapid and sensitive methods of detecting STIs in order for patients to get results and treatment immediately. It is important that patients receive results and initial consultation or treatment during the first patient visit, as they rarely return for a second visit. Point-of-care (POC) tests are an important strategy to address the STI epidemic, and because many affected by STIs (young and/or poor) often have limited access to traditional healthcare, POC tests that can be performed in non-traditional settings (e.g., university clinics, community public health care clinics, jails, and detention centers) could greatly reduce STIs. The system we propose to further develop will combine AI Biosciences, Inc.’s novel assay cartridge, low-cost automatic platform inspired by 3D printer technology, and a rapid real-time thermal cycler to form a multi- sample system for use in non-traditional healthcare settings. A low-cost self-contained nucleic acid extraction and purification cartridge will be developed to perform a 10 min. extraction process for 6 samples. The extracted NA samples will then be eluted and amplified using shuttle thermal cycler to perform multiplexed real-time polymerase chain reaction (PCR) for CT and NG targets (15 min for 40 cycles). We plan to optimize the assay and disposable to meet the needs of non-traditional healthcare settings, improve the system for ease-of-use, and minimize risk of erroneous result. We will complete prototype system validation and verification as well as determine analytical sensitivity and specificity of the CT/NG assay. The proposed work will be carried out in collaboration with a STI expert in the US. Our proposed technologies are highly suitable for parallel, automated processing. This is a substantial advantage over other integrated systems being developed as most can only process one sample at a time, requiring the purchase of several modules in order to meet the throughput demand in most point-of-care or near-patient settings. The successful development and commercialization of our medium-throughput technology will impact how molecular diagnostic assays are implemented in non-traditional healthcare settings.