Rapid Multiplex RT-PCR for Influenza A and B by Genesoc^®, a Microfluidic PCR System

Area of Science:

• Molecular Diagnostics
• Virology
• Microfluidics

Background:

• Rapid antigen tests are common for influenza diagnosis but have low sensitivity.
• There is a need for more sensitive molecular diagnostic tests for influenza A and B.
• Microfluidic thermal cycling technology offers potential for rapid molecular testing.

Purpose of the Study:

• To develop and clinically evaluate a rapid multiplex testing protocol for influenza A and B using the GeneSoC® microfluidic real-time PCR system.
• To compare the performance of the GeneSoC® assay against conventional real-time RT-PCR and rapid antigen tests.

Main Methods:

• Developed a rapid real-time PCR protocol on the GeneSoC® microfluidic system for influenza A and B detection.
• Validated specificity with influenza A/B, human metapneumovirus, and RSV strains.
• Evaluated analytical sensitivity using synthesized RNA and clinical nasopharyngeal samples.
• Performed cross-validation against conventional RT-PCR and rapid antigen tests.

Main Results:

• The GeneSoC® assay detected influenza A and B at low concentrations (38 and 65 copies/µL).
• 100% agreement was observed between GeneSoC® RT-PCR and conventional RT-PCR for clinical specimens.
• GeneSoC® RT-PCR showed 95.7% overall agreement with rapid antigen tests.
• The average turnaround time for GeneSoC® RT-PCR was approximately 16.5 minutes.

Conclusions:

• The GeneSoC® microfluidic real-time PCR system demonstrates analytical performance comparable to conventional RT-PCR.
• The system offers a rapid turnover time, making it a viable alternative to current rapid antigen tests for influenza diagnosis.