Development of a multiplex and universal RT-PCR assay for hemagglutinin and neuraminidase subtyping of avian influenza virus.

ABSTRACT:

Importance

Avian influenza virus (AIV) is classified into subtypes by hemagglutinin (HA) and neuraminidase (NA) genes. Wild waterfowl harbor H1-H16 and N1-N9 subtypes. Large-scale AIV surveillance requires substantial labor, time, and cost when using conventional HA and NA subtyping methods.

Objective

This study aimed to develop multiplex reverse transcription polymerase chain reaction (RT-PCR) and universal assays to detect all H1-H15 and N1-N9 subtypes in just eight RT-PCR reactions.

Methods

Subtype-specific forward primers and conserved reverse primers were used to identify HA and NA subtypes based on RT-PCR amplicon size. The HA cleavage site was characterized by sequencing. For NA, a universal forward primer enabled sequencing-based subtyping from a single reacti

SUBMITTER: An SH 

PROVIDER: S-EPMC12511891 | biostudies-literature | 2025 Sep

REPOSITORIES: biostudies-literature