Detection of influenza A virus subtypes using a solid-phase PCR microplate chip assay

• We have developed a microplate chip microarray based solid-phase PCR.
• The microplate chip was produced in the microplate not on the glass slides.
• Ezymatic colorimetric and fluorescence methods were developed for signal detection.
• It can rapidly detect general influenza virus type A and identified 9 subtypes.
• The platform can be also used in the pathogenic microorganism detection.

A rapid and sensitive microplate chip based on solid PCR was developed to identify influenza A subtypes. A simple ultraviolet cross-linking method was used to immobilize DNA probes on pretreated microplates. Solid-phase PCR was proven to be a convenient method for influenza A screening. The sensitivity of the microplate chip was 10−3 μg/mL for the enzymatic colorimetric method and 10−4 μg/mL for the fluorescence method. The 10 sets of primers and probes for the microplate chip were highly specific and did not interfere with each other. These results suggest that the microplate chip based on solid PCR can be used to rapidly detect universal influenza A and its subtypes. This platform can also be used to detect other pathogenic microorganisms.