A novel sample processing method for the co-extraction of DNA/RNA from Mycobacterium tuberculosis (TB) and SARS-COV-2, complementing a novel near-patient CRISPR test for tuberculosis detection (CRISPR SHINE TB).
Invention Summary:
Tuberculosis and COVID-19 are infectious diseases with high mortality rates caused by Mycobacterium tuberculosis (TB) and SARS-CoV-2. Co-infection is increasing in prevalence, but methods for single-sample co-detection are limited by efficacy and ease of use. As the areas with particularly high rates of co-infection worldwide are often limited in laboratory and clinical capabilities, there is a need to develop simple and accurate sample preparation and testing methods. Current extraction methods of DNA and RNA from the same sample often involve complex processes and expensive or hazardous reagents and have limited downstream applications. Additionally, while tests do exist for TB, simple and accurate kits with sample processing and detection reagents are not currently available.
Rutgers and Princeton researchers and their collaborators have invented: (1) an easy and efficient method for co-extraction of TB DNA and COVID-19 RNA that is compatible with many downstream assays, including qPCR, PCR, and CRISPR, and (2) a CRISPR diagnostic tool for TB with point-of-care capabilities. The extraction method utilizes a simple Chelex resin-based boiling method and is effective on both saliva and sputum samples. The CRISPR SHINE-TB diagnostic uses two single-pot reactions with isothermal amplification by recombinase polymerase amplification (RPA), in vitro transcription, and detection, using Cas13a for TB detection and Cas12a for internal control detection. Point-of-care optimization is ongoing. This invention enables sample processing and testing for COVID-19 and TB from the same sample, which is advantageous for individuals at high risk of co-infection.
Market Applications:
Efficient co-extraction of DNA and RNA from a single sample.
Simple nucleic acid extraction requiring minimal small, low-cost equipment.
Compatible extraction reagents for downstream use with various molecular assays (e.g., PCR, isothermal amplification).
Near patient TB diagnostic kits (CRISPR SHINE-TB) offering rapid, sensitive, and specific detection outside traditional labs.
Integrated test kits combining extraction and diagnostic components for streamlined TB/COVID co-detection at clinical or remote sites.
Advantages:
Simplified protocol requiring minimal equipment and technical expertise.
Cost-effective compared to existing complex and expensive extraction kits.
Dual-pathogen extraction for simultaneous isolation of TB DNA and SARS-CoV-2 RNA from a single sample, enabling efficient multiplex testing in labs and clinics.
Broad downstream compatibility with various molecular assays.
Robust sensitivity and specificity of CRISPR SHINE-TB, compared to leading TB tests.
Portable and user-friendly, enabling use in low-resource settings.
Publications:
Dunkley ORS, Bell AG, Modi NH, Huang Y, Tseng S, Reiss R, Daivaa N, Davis JL, Vargas DA, Banada P, Xie YL, Myhrvold C. A Streamlined Point-of-Care CRISPR Test for Tuberculosis Detection Directly from Sputum. medRxiv [Preprint]. 2025 Mar 4:2025.02.19.25322517. doi: 10.1101/2025.02.19.25322517. PMID: 40034782; PMCID: PMC11875272.
Modi NH, Dunkley ORS, Bell AG, Hennig E, Wats A, Huang Y, Daivaa N, Myhrvold C, Xie YL, Banada P. Simplified Co-extraction of total Nucleic Acids from Respiratory Samples for detection of Mycobacterium tuberculosis and SARS-CoV-2 optimized for compatibility across Diagnostic Platforms. medRxiv [Preprint]. 2025 Mar 3:2025.02.27.25322880. doi: 10.1101/2025.02.27.25322880. PMID: 40093238; PMCID: PMC11908299.
Intellectual Property & Development Status: Provisional application filed. Patent pending. Available for licensing and/or research collaboration. For any business development and other collaborative partnerships, contact: marketingbd@research.rutgers.edu