Preliminary study of specific purification and enrichment method for viral genome based on nucleic acid hybridization
10.3760/cma.j.issn.1003-9279.2016.02.025
- VernacularTitle:基于核酸杂交的病毒基因组特异性纯化富集方法的初步研究
- Author:
Lijing WANG
1
;
Lin LIU
;
Jiandong LI
;
Weihong LI
;
Shou ZHANG
;
Mifang LIANG
;
Dexin LI
Author Information
1. 中国疾病预防控制中心病毒病预防控制所
- Keywords:
Viral genome;
Enrichment;
Purification;
Next-Generation sequencing
- From:
Chinese Journal of Experimental and Clinical Virology
2016;30(2):212-215
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the specific purification and enrichment method for the viral genome in clinical samples before high-throughput detection.Methods The negative chain RNA virus fever with thrombocytopenia syndrome (SFTSV) as was taken the simulate samples and the hybridization probes of the segment S,M and L were desiqned respectively,modified the probes with biotin in 5 ' ends to covalent bonded with streptavidin magnetic beads to purify the viral genome and compared the influence of specific purification on the effect of high-throughput detection.Results Specific enrichment and purification for viral genome before sequencing can significantly increase the proportion of valid data and whole genome coverage.The valid data proportions of the groups purified before or after whole transcriptome amplification (WTA) were improved significantly compared to group without purifying.The valid data proportion respectively were 5.57％,6.39％,2.71％,and the difference was significant((x2 =9799.8,P ＜0.001)).And the effect of the group purified after whole transcriptome amplification was better than group purified before whole transcriptome amplification(P ＜ 0.001).Conclusions Viral genome specific enrichment and purification method based on nucleic acid hybridization can effectively reduce the background values in the second generation sequencing,improve the sequencing efficiency and play the role of purification and enriching the nucleic acid in samples specifically before high-throughput detection.
