A Novel Sample-to-Answer Visual Nucleic Acid Detection System for Adenovirus Detection.

Area of Science:

• Molecular Biology
• Point-of-Care Diagnostics
• Virology

Background:

• Human adenoviruses (HAdVs) cause outbreaks and public health threats.
• Rapid, accessible diagnostics are crucial for controlling HAdV spread, especially in resource-limited areas.
• Existing methods often require specialized equipment and trained personnel, limiting field applicability.

Purpose of the Study:

• To develop an integrated, electricity-independent sample-to-answer system for HAdV detection.
• To enable rapid, sensitive, and specific HAdV diagnosis in resource-limited and field settings.
• To create a portable and disposable detection system for AdvB and AdvE.

Main Methods:

• Developed a two-module system: ALP FINA (alkaline lysis with paper-based filtration) for nucleic acid extraction and SV RPA (sealed and visual recombinase polymerase amplification) for detection.
• The system operates at room temperature, requiring no external electricity or complex laboratory infrastructure.
• Tested on nasopharyngeal swab samples from patients infected with AdvB/AdvE and healthy volunteers.

Main Results:

• ALP FINA achieved nucleic acid extraction efficiency of 48-84%, comparable to conventional methods.
• SV RPA demonstrated high sensitivity (close to 10 copies/μL) and prevented aerosol contamination.
• The complete system achieved 100% sensitivity and 100% specificity in detecting AdvB and AdvE in clinical samples.

Conclusions:

• The developed portable, disposable, and modular detection system is effective for AdvB and AdvE.
• Its electricity-independent nature makes it ideal for resource-limited settings and field applications.
• This system holds significant potential for early and rapid diagnosis of HAdV infections in diverse environments.