Nanomicroarray and multiplex next-generation sequencing for simultaneous identification and characterization of influenza viruses

Abstract

Conventional methods for detection and discrimination of infuenza viruses are time consuming and labor inten sive. We developed a diagnostic platform for simultaneous identifcation and characterization of infuenza viruses that uses a combination of nanomicroarray for screening and multiplex next-generation sequencing (NGS) assays for laboratory confrmation. The nanomicroarray was devel oped to target hemagglutinin, neuraminidase, and matrix genes to identify infuenza A and B viruses. PCR ampli- cons synthesized by using an adapted universal primer for all 8 gene segments of 9 infuenza A subtypes were detected in the nanomicroarray and confrmed by the NGS assays. This platform can simultaneously detect and dif ferentiate multiple infuenza A subtypes in a single sample. Use of these methods as part of a new diagnostic algo rithm for detection and confrmation of infuenza infections may provide ongoing public health benefts by assisting with future epidemiologic studies and improving prepared- ness for potential infuenza pandemics.