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Unique Barcoded Primer-Assisted Sample-Specific Pooled Testing (Uni-Pool) for Large-Scale Screening of Viral
Xinyu Zhuang1, Xiao Lu1, Henson L Lee Yu1
1Department of Chemical and Biological Engineering, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong 999077, China.
Analytical Chemistry
|February 24, 2022
Summary
Pooled testing for COVID-19 can be improved with Uni-Pool, a new strategy that uses barcoded primers to avoid retesting. This method accurately identifies positive and negative samples in a pool, saving time and resources during high-prevalence periods.
Area of Science:
- Molecular Biology
- Virology
- Biotechnology
Background:
- Pooled testing is crucial for large-scale COVID-19 screening, offering cost and time savings.
- A major drawback of pooled testing is the need for retesting individual samples when a pool tests positive, especially during high infection waves.
- Existing methods face challenges in maintaining accuracy and sensitivity due to sample dilution in pooled formats.
Purpose of the Study:
- To develop a novel pooled testing strategy, Uni-Pool, that eliminates the need for retesting.
- To enhance the efficiency of large-scale infectious disease screening.
- To maintain high specificity and sensitivity in pooled testing without compromising detection limits.
Main Methods:
- Developed a unique barcoded primer-assisted sample-specific pooled testing strategy (Uni-Pool).
- Extracted and amplified key viral genetic sequences with sample-specific identifiers.
- Utilized multiplex melting curve analysis post-real-time polymerase chain reaction for result determination.
Main Results:
- Uni-Pool successfully differentiated positive and negative samples within a single pool with negligible cross-reactivity.
- Demonstrated the method's efficacy with 40 simulated samples containing SARS-CoV-2 pseudovirus across various loads.
- Validated the successful testing of five samples in one pool with a 15% infection prevalence.
Conclusions:
- Uni-Pool significantly reduces assay time for large-scale screening by eliminating retesting steps.
- The strategy maintains specificity and detection sensitivity, overcoming limitations of traditional pooled testing.
- Uni-Pool offers a robust and efficient solution for infectious disease surveillance, particularly during pandemics.
