AHS further characterization: Third step
Genome sequencing could reveal important information about the origins, evolution, and characteristics of each viral population (e.g., reassortment, pathogenicity). Until recent, Sanger sequencing based on “conserved primers” has been the principal sequencing methodology for BTV and is still used nowadays in epidemiological studies, specially to seg-2, since it is the most variable region.
Cloning/PCR amplification of dsRNA genome segments sequence-independent approaches were developed. These include the single primer amplification technique or SPAT (Attoui et al., 2000; Lambden et al., 1992),and a derived more effective version, the full-length amplification of complementary DNA or FLAC (Maan et al., 2007b). These methods allow sequence determination of individual full-length genome segments or even full-length genomes.
However, NGS approaches are increasingly becoming the methods of choice for sequencing RNA virus genomes, including BTV. Several NGS technologies have been used for BTV genome sequence characterizations. (Guthrie et al., 2015; King et al., 2020; Toh et al., 2020). The consequences of these continuously evolving methodologies for sequence determination have resulted in an immense expansion of the BTV sequence databases.
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