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Related Experiment Video

Updated: Dec 23, 2025

Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a

Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a

Published on: December 23, 2022

Massively multiplexed nucleic acid detection with Cas13.

Cheri M Ackerman^1,2, Cameron Myhrvold^3,4, Sri Gowtham Thakku^1,5

¹Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, MA, USA.

|April 30, 2020

Summary

This summary is machine-generated.

A new platform, Combinatorial Arrayed Reactions for Multiplexed Evaluation of Nucleic acids (CARMEN), enables scalable, multiplexed pathogen detection using CRISPR technology. This advancement improves disease surveillance and diagnostics by allowing simultaneous testing of thousands of targets in numerous samples.

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Area of Science:

Molecular Biology
Biotechnology
Infectious Disease Surveillance

Background:

Current pathogen detection methods struggle with scalability and multiplexing, limiting their effectiveness in routine surveillance and outbreak response.
There is a critical need for advanced diagnostic technologies capable of testing numerous samples for a wide range of pathogens simultaneously.

Purpose of the Study:

To develop a scalable and highly multiplexed platform for nucleic acid detection.
To enhance pathogen surveillance and diagnostic capabilities for global health security.

Main Methods:

Development of the Combinatorial Arrayed Reactions for Multiplexed Evaluation of Nucleic acids (CARMEN) platform.
Integration of CARMEN with CRISPR-Cas13 detection for high-throughput nucleic acid analysis.
Utilizing nanolitre droplets in microwell arrays for sample-reagent pairing and replicate testing.

Main Results:

The CARMEN-Cas13 platform enables robust testing of over 4,500 CRISPR RNA-target pairs on a single array.
Successfully developed a multiplexed assay capable of differentiating all 169 human-associated viruses and detecting SARS-CoV-2.
Demonstrated comprehensive subtyping of influenza A strains and identification of HIV drug-resistance mutations.
Achieved over 300-fold reduction in reagent cost per test through miniaturization and scalability.

Conclusions:

CARMEN-Cas13 offers a scalable, cost-effective solution for highly multiplexed nucleic acid detection.
This technology significantly advances capabilities for both clinical diagnostics and large-scale public health surveillance.
The platform facilitates a shift towards comprehensive testing of large sample sets, improving patient care and outbreak preparedness.