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Principal Investigator: Peter B Lillehoj
Organization: RICE UNIVERSITY
Fiscal Year: 2024
Award: $449,888
Funding agency: National Institute of Allergy and Infectious Diseases
HIV is one of the most deadly infectious diseases in the world. HIV transmission is largely driven by acutely
infected individuals who are unaware of their status or those receiving antiretroviral therapy but not virally
suppressed. Currently available rapid HIV tests, including fourth-generation HIV antigen/antibody combo tests,
lack the sensitivity needed for detecting HIV during early stages of acute infection and viral rebound. Nucleic
acid amplification tests (NAATs) that detect HIV-1 RNA, which can be detected in as early as 5 to 10 days after
viral transmission, offer superior sensitivity and are currently the gold standard for detecting HIV during acute
infection and viral rebound. However, NAATs are laborious, time-consuming and rely on expensive
instrumentation and refrigerated components, hindering their use for self-testing.
Therefore, the objective of this project is to develop and validate a shelf-stable, instrument-free CRISPR-
Cas13-based rapid test for highly sensitive HIV-1 detection in blood or saliva. This high-risk, high-reward
approach is fundamentally different from existing NAATs that involve multiple steps of sample preparation (RNA
extraction and amplification) or require laboratory instrumentation or refrigerated components. Our rapid HIV-1
test will employ an engineered Cas13a enzyme with enhanced collateral activity and stability, and lyophilized
assay components for enhanced shelf life while offering a simplified workflow that can be performed in home
settings (similar to COVID-19 antigen rapid self-tests). A phone app will automatically analyze the readout from
the lateral flow device and present the test results in a clear, simple-to-interpret manner. The rationale for this
research is supported by the applicants' preliminary data demonstrating highly sensitive detection of viral RNA
using a rapid, lateral flow CRISPR-Cas13-based assay and the development of an RRM2-L Cas13a enzyme
with enhanced collateral activity and stability. In the R61 phase, we will pursue the following specific aims:
1) Characterize HIV-1 RNA levels in saliva and blood from a diverse population of people with HIV;
2) Investigate strategies for user-friendly device operation;
3) Develop a shelf-stable, instrument-free CRISPR-Cas13 rapid test for highly sensitive HIV-1 detection.
If the R61 phase milestones are achieved, we will further develop and characterize this platform and validate its
functionality using clinical HIV specimens during the R33 phase by pursuing the following specific aims:
4) Further develop and characterize the lateral flow CRISPR-Cas13-based rapid HIV-1 test;
5) Validate the functionality of the CRISPR-Cas13-based rapid HIV-1 test using clinical specimens.
This approach is innovative because it combines the speed, simplicity and affordability of lateral flow-based
diagnostic tests with the high sensitivity of CRISPR-Cas-based nucleic acid detection, and it is significant
because it will improve the effectiveness of HIV self-testing for early diagnosis and patient monitoring during ART
therapy, thus minimizing unintentional HIV transmission and reducing HIV-associated morbidity and mortality.
Terms: <AIDS Virus><AIDS test><AIDS/HIV test><Acquired Immune Deficiency Syndrome Virus><Acquired Immunodeficiency Syndrome Virus><Acute><Android App><Android Application><Antibodies><Architecture><Assay><Bathing><Baths><Bioassay><Biological Assay><Blood><Blood Reticuloendothelial System><Blood Sample><Blood specimen><COVID-19 antigen><CRISPR><CRISPR/Cas system><Cell Phone Application><Cell phone App><Cellular Phone App><Cellular Phone Application><Chronic><Clinical><Clinical Management><Clustered Regularly Interspaced Short Palindromic Repeats><Communicable Diseases><Consumption><Data><Detection><Development><Devices><Diagnosis><Diagnostic><Diagnostic tests><Early Diagnosis><Effectiveness><Engineering><Engineering / Architecture><Enzyme Gene><Enzymes><Evaluation><FDA approved><Freeze Drying><Freeze Dryings><General Population><General Public><Generations><HIV><HIV Antigens><HIV Infections><HIV test><HIV-1><HIV-1 test><HIV-2 test><HIV-Associated Antigens><HIV-I><HIV1><HTLV-III Antigens><HTLV-III Infections><HTLV-III-LAV Antigens><HTLV-III-LAV Infections><Hand><Health><Home><Human><Human Immunodeficiency Virus Type 1><Human Immunodeficiency Viruses><Human T-Lymphotropic Virus Type III Antigens><Human T-Lymphotropic Virus Type III Infections><Human immunodeficiency virus 1><Human immunodeficiency virus test><Hydrogen Oxide><Individual><Infectious Disease Pathway><Infectious Diseases><Infectious Disorder><LAV Antigens><LAV-HTLV-III><Laboratories><Lateral><Life><Lymphadenopathy-Associated Antigens><Lymphadenopathy-Associated Virus><Lyophilization><Measurement><Methods><Mission><Modern Man><Morbidity><Morbidity - disease rate><NIAID><National Institute of Allergy and Infectious Disease><Non-Polyadenylated RNA><Nucleic Acid Amplification Tests><Nucleic Acid Testing><Nucleic Acids><Outcome><Patient Monitoring><Patients><Performance><Persons><Phase><Phone><Population Heterogeneity><PrEP><Preparation><Procedures><Public Health><R2 Gene><RNA><RNA Gene Products><RNA amplification><RR2 Gene><RRM2><RRM2 gene><Reaction><Refrigeration><Research><Research Specimen><Ribonucleic Acid><Ribonucleoside-Diphosphate Reductase M2 Chain Gene><Ribonucleotide Reductase M2 Polypeptide Gene><Ribonucleotide Reductase Small Chain Gene><SARS-CoV-2 antigen><Saliva><Sampling><Sensitivity and Specificity><Smart Phone App><Smart Phone Application><Smartphone App><Specimen><Speed><Techniques><Technology><Telephone><Temperature><Test Result><Testing><Time><Transmission><Viral Gene Products><Viral Gene Proteins><Viral Proteins><Virus-HIV><Water><acute infection><antiretroviral therapy><antiretroviral treatment><assess effectiveness><cell phone based app><comparative><coronavirus disease 2019 antigen><cost><determine effectiveness><developmental><diagnostic ability><diagnostic capability><diagnostic power><diagnostic tool><diagnostic utility><diagnostic value><diverse populations><early detection><effectiveness assessment><effectiveness evaluation><evaluate effectiveness><examine effectiveness><hands><heterogeneous population><high reward><high risk><homes><iOS app><iOS application><iPhone App><iPhone Application><improved><innovate><innovation><innovative><instrument><instrumentation><isothermal amplification><lateral flow assay><lateral flow test><lateral line><mobile phone app><mortality><new diagnostics><next generation diagnostics><novel diagnostics><nucleic acid detection><operation><operations><population diversity><pre-exposure prophylaxis><preparations><prototype><rapid assay><rapid test><rapid tests><saliva assay><saliva based assay><saliva based test><saliva test><salivary assay><salivary test><self testing><severe acute respiratory syndrome coronavirus 2 antigen><smartphone application><smartphone based app><smartphone based application><test strip><transmission process><usability><user-friendly><viral RNA><viral detection><viral rebound><viral transmission><virus RNA><virus detection><virus protein><virus rebound><virus transmission>