A Multiplex CRISPR-Cas12a-Based Hydrogel Microarray Platform for the Simultaneous Detection of Common Adenovirus...

Area of Science:

• Molecular Biology
• Biotechnology
• Infectious Diseases

Background:

• Adenovirus infections pose a significant global health burden, causing respiratory and gastrointestinal diseases.
• Current diagnostic methods like PCR are slow and require extensive lab facilities, hindering use in resource-limited settings.
• Accurate and rapid adenovirus typing is crucial for effective patient management and disease control.

Purpose of the Study:

• To develop a novel CRISPR/Cas12a-based assay for simultaneous detection of six common adenovirus types.
• To integrate the assay with a hydrogel microarray for enhanced efficiency and multiplexing.
• To evaluate the sensitivity, specificity, and clinical applicability of the developed diagnostic platform.

Main Methods:

• Utilized a CRISPR/Cas12a system combined with recombinase polymerase amplification (RPA) for DNA amplification.
• Developed a hydrogel microarray to immobilize Cas12a-crRNA complexes and capture amplified viral DNA.
• Generated a fluorescent signal upon Cas12a activation, indicating the presence of specific adenovirus types.

Main Results:

• The assay successfully detected six common adenovirus types (1, 2, 3, 4, 7, and 14) simultaneously.
• Achieved high sensitivity with detection limits between 10-50 copies/μL for all targeted types.
• Demonstrated excellent specificity, showing no cross-reactivity with other respiratory viruses and 100% specificity in clinical validation.

Conclusions:

• The CRISPR/Cas12a hydrogel microarray platform provides a rapid, cost-effective, and highly specific method for adenovirus detection.
• This innovative diagnostic tool has significant potential for improving clinical diagnostics and public health surveillance.
• The assay offers a promising alternative to traditional methods, especially in settings with limited resources.