Establishment and evaluation of a genome-wide multiplex PCR sequencing method for severe fever with thrombocytopenia syndrome virus based on high-throughput sequencing technology
10.3760/cma.j.cn112309-20220611-00196
- VernacularTitle:基于高通量测序技术的发热伴血小板减少综合征病毒全基因组多重PCR测序方法的建立与评价
- Author:
Xiaoyang WANG
1
;
Shujun PEI
;
Zeqian CHEN
;
Wenshuai SUO
;
Zhijie YI
;
Yi LI
;
Xueyong HUANG
;
Wanshen GUO
Author Information
1. 郑州大学公共卫生学院流行病学教研室，郑州 450001
- Keywords:
Severe fever with thrombocytopenia syndrome virus;
Multiplex PCR;
High-throughput sequencing;
Genome
- From:
Chinese Journal of Microbiology and Immunology
2022;42(12):931-939
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a sequencing method for the genome of severe fever with thrombocytopenia syndrome virus (SFTSV) based on next-generation sequencing (NGS).Methods:SFTSV RNA was extracted from serum samples of patients with severe fever with thrombocytopenia syndrome. SFTSV-specific primers were designed using Primer 5.0 software. A multiplex PCR method was constructed and used to amplify the nucleotide sequence of SFTSV. Whole-genome sequencing was performed on the NGS platform.Results:The whole genes of SFTSV isolates in 28 serum samples were amplified by the multiplex PCR with a coverage over 94%. Sequencing and phylogenetic analysis of those strains revealed that the predominant strains ( n=20) belonged to genotype A, followed by genotypes B ( n=4) and E ( n=3). Conclusions:A high-throughput sequencing method for SFTSV based on multiplex PCR was established in this study. This method was characterized by high specificity and good quality and could improve the sequencing efficiency.
