Establishment of a high throughput sequencing technology for detection of SFTS virus genome
10.3760/cma.j.issn.1003-9279.2019.01.020
- VernacularTitle: 基于高通量技术的SFTS病毒基因组测序方法的建立
- Author:
Weiwei YU
1
;
Yi SUN
2
;
Hao YAN
2
;
Wenwu YAO
2
;
Xiuyu LOU
2
;
Haiyan MAO
2
;
Yanjun ZHANG
2
Author Information
1. College of Medical Technology, Zhejiang Chinese Medical University, Hangzhou 310053, China
2. Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310051, China
- Publication Type:Journal Article
- Keywords:
Severe fever with thrombocytopenia syndrome virus;
Library construction;
Next-generation sequencing;
Genome
- From:
Chinese Journal of Experimental and Clinical Virology
2019;33(1):89-94
- CountryChina
- Language:Chinese
-
Abstract:
Objective:The objective of this study was to establish a next generation sequencing (NGS) method for severe fever with thrombocytopenia syndrome virus(SFTS).
Methods:SFTS virus RNA was extracted from the patient serum inoculated and isolated by Vero cells. Two methods of random primer sequencing and oligo(dT) beads selection sequencing were used for library construction. The libraries were built based on the best amplification and purification conditions. Whole genome sequencing was performed on NGS platform.
Results:There were significant differences in data of 3 virus between the two methods.The sample was sequenced by random primer sequencing showed low coverage and depth. However, three samples were sequenced by oligo(dT) beads selection showed coverage was over 99% and depth was over 900.The alignment rate of database from NCBI was more than 90%. The initial detection quality of this method was 300ng RNA.
Conclusions:In this study, we used the method of oligo(dT) beads selection to build libraries, and established a SFTS virus genome detection based on next generation sequencing.
