Culture-Independent Workflow for Nanopore MinION-Based Sequencing of Influenza A Virus
Mojnu Miah, Mohammad Enayet Hossain, Rashedul Hasan, Md. Shaheen Alam, Joynob Akter Puspo, Md. Mahmudul Hasan, Ariful Islam, Sukanta Chowdhury, Mohammed Ziaur Rahman
Abstract
The Nanopore MinION-based influenza sequencing approach we are proposing makes it possible to sequence the influenza A virus, irrespective of its diverse serotypes, directly from clinical and environmental swab samples, so that we are not limited to virus culture. This third-generation, portable, multiplexing, and real-time sequencing strategy is highly convenient for local sequencing, particularly in low- and middle-income countries like Bangladesh. Furthermore, the cost-efficient sequencing method could provide new opportunities to respond to the early phase of an influenza pandemic and enable the timely detection of the emerging subtypes in clinical samples. Here, we meticulously described the entire process that might help the researcher who will follow this methodology in the future. Our findings suggest that this proposed method is ideal for clinical and academic settings and will aid in real-time surveillance and in the detection of potential outbreak agents and newly evolved viruses.