Next-generation sequencing library preparation method for identification of RNA viruses on the Ion Torrent Sequencing Platform
Next generation sequencing (NGS) is a powerful tool for the characterization, discovery, and molecular identification of RNA viruses. There were multiple NGS library preparation methods published for strand-specific RNA-seq, but some methods are not suitable for identifying and characterizing RNA viruses. In this study, we report a NGS library preparation method to identify RNA viruses using the Ion Torrent PGM platform. The NGS sequencing adapters were directly inserted into the sequencing library through reverse transcription and polymerase chain reaction, without fragmentation and ligation of nucleic acids. The results show that this method is simple to perform, able to identify multiple species of RNA viruses in clinical samples.
KeywordsNext-generation sequencing Library preparation RNA virus
This study was funded by the National Key Research and Development Program of China (2017YFC120050) and China Animal Health and Epidemiology Center Innovation Fund (2015IF-0004FF).
KW supervised the research, contributed to the design and development of the experimental work, and wrote the paper. GC contributed to the experimental work and the bioinformatics analysis. YQ and QZ contributed to the experimental work. SW performed the revise of the language. TW performed the bioinformatics analysis.
Compliance with ethical standards
Conflict of interest
Kaicheng Wang, Guiqian Chen, Yuan Qiu, Qingye Zhuang, Suchun Wang, and Tong Wang have received research grants from China Animal Health and Epidemiology Center. All the authors declare that they have no conflict of interest.
Informed consent was obtained from all individual participants included in the study.
Research involving human participants and/or animals
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.
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