Capillarity-powered and CRISPR/Cas12a-responsive DNA hydrogel distance sensor for highly sensitive visual detection of HPV DNA

Aug 13, 2024Biosensors & bioelectronics

Highly sensitive visual detection of HPV DNA using a capillary-powered DNA hydrogel sensor activated by CRISPR/Cas12a

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Abstract

The sensor can detect as low as 1 aM HPV 16 DNA within 30 minutes.

The DNA hydrogel distance sensor utilizes capillarity and CRISPR/Cas12a technology for point-of-care testing.
Target DNA activates the Cas12a enzyme, which cleaves crosslinked DNA in the hydrogel, increasing its permeability.
The presence of target DNA allows a sample solution to travel further in the capillary tube compared to negative samples.
This design enables rapid and sensitive detection of HPV infection without the need for special equipment.
The sensor's features include high sensitivity, specificity, and a user-friendly read-out.

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The rapid and specific identification and sensitive detection of human papillomavirus (HPV) infection is critical for preventing cervical cancer, particularly in resource-limited regions. In this work, we hope to propose a capillarity-powered and CRISPR/Cas12a-responsive DNA hydrogel distance sensor for point-of-care (POC) DNA testing. Using the thermal reversibility of DNA hydrogel and capillarity, the novel DNA hydrogel distance sensor can be rapidly and simply constructed by loading an ultra-thin CRISPR/Cas12a-responsive DNA-crosslinked hydrogel film at the end of the capillary tube. The target DNA-specific recombinase polymerase reaction (RPA) amplicons activate the trans-cleavage activity of the Cas12a enzyme, cleaving the crosslinked DNA in hydrogel film, and causing an increase of hydrogel's permeability. As a result, a sample solution containing target DNA travels into the capillary tube at a longer distance compared to the negative samples. Reading the solution traveling distance in capillary tubes, the novel sensor realizes target DNA detection without any special equipment. Benefiting from the exponential target amplification of RPA and multiple turnover response of trans-cleavage of CRISPR/Cas12a, the developed sensor can visually and specifically detect as low as 1 aM HPV 16 DNA within 30 min. These outstanding features, including exceptional sensitivity and specificity, simple and portable design, mild measurement conditions, quick turnaround time, and user-friendly read-out, make the novel distance sensor a promising option for POC diagnostic applications.

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Capillarity-powered and CRISPR/Cas12a-responsive DNA hydrogel distance sensor for highly sensitive visual detection of HPV DNA

Abstract

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