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

Author:

Luo Junhong12ORCID,Huang Yao3,Guo Yan4,Meng Zixinrong2,Zhao Kangchen1,Zhu Xiaojuan1,Zhou Le3,Wu Tao1,Qiao Qiao1,Ge Yiyue1,Xu Qin3,Cui Lunbiao12

Affiliation:

1. NHC Key laboratory of Enteric Pathogenic Microbiology, Jiangsu Province Engineering Research Center of Health Emergency, Jiangsu Provincial Center for Disease Control & Prevention, Nanjing, China

2. School of Public Health, Nanjing medical university, Nanjing, China

3. Yangzhou City Center for Disease Control & Prevention, Yangzhou, China

4. Department of Neonatology, Children's Hospital of Nanjing Medical University, Nanjing, China

Abstract

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.

Funder

Yangzhou Science and Technology Planning Project

Jiangsu Provincial Health Commission scientific research project

Natural Science Foundation of Jiangsu Province

Publisher

Future Medicine Ltd

Subject

Virology

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