Nanopore sequencing approach for variation detection of a live attenuated Newcastle disease virus vaccine.

Live attenuated Newcastle disease virus (NDV) vaccines have been used widely to protect chickens against Newcastle disease. However, the vaccine issues caused by genome mutations can seriously affect poultry health. In this study, the authors demonstrate the use of nanopore sequencing technology for rapid genome determination and variation analysis from a live attenuated NDV vaccine. NDV-specific reads were detected immediately after sequencing, and 24× genome coverage was obtained within 10 min. Variation analysis revealed 19 variant sites across the vaccine genome compared to the NDV clone 30 reference sequence . The sequencing and data analysis workflow employed enables all basic molecular biology laboratories to perform detailed genome sequencing in live attenuated vaccine, providing an effective means of quality control for vaccine production.