Establishment and evaluation of a multiplex PCR amplification-based sequencing method for respiratory virus type A

Aim: This study aimed to establish a simple whole-genome sequencing method for respiratory virus type A (RSVA) used in genomic epidemiological studies. Methods: Multiple primers were designed to amplify overlapping amplicons specific to RSVA. The amplicons were sequenced using Illumina and Nanopore sequencing platforms and the sequencing performances of the two platforms were also compared. Results: The study shows that the developed method could recover almost full genomes of RSVA when Ct values were less than 33 for clinical samples. Conclusion: In this study, a whole-genome sequencing method was developed for RSVA based on multiplex PCR.