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Emerg Infect Dis. 2015 Mar;21(3):400-8. doi: 10.3201/eid2103.141169.

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

Abstract

Conventional methods for detection and discrimination of influenza viruses are time consuming and labor intensive. We developed a diagnostic platform for simultaneous identification and characterization of influenza viruses that uses a combination of nanomicroarray for screening and multiplex next-generation sequencing (NGS) assays for laboratory confirmation. The nanomicroarray was developed to target hemagglutinin, neuraminidase, and matrix genes to identify influenza A and B viruses. PCR amplicons synthesized by using an adapted universal primer for all 8 gene segments of 9 influenza A subtypes were detected in the nanomicroarray and confirmed by the NGS assays. This platform can simultaneously detect and differentiate multiple influenza A subtypes in a single sample. Use of these methods as part of a new diagnostic algorithm for detection and confirmation of influenza infections may provide ongoing public health benefits by assisting with future epidemiologic studies and improving preparedness for potential influenza pandemics.

PMID:
25694248
PMCID:
PMC4344273
DOI:
10.3201/eid2103.141169
[Indexed for MEDLINE]
Free PMC Article
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