Genotyping targeting the capsid genes was conducted by previously described PCR and sequencing methods [7 (link)8 (link)]. Briefly, rotavirus G (VP7) and P (VP4) genotyping was performed according to the WHO manual, using specific primer sets [9 ], with some modifications. Norovirus capsid genotyping was performed using specific primer sets [8 (link)], and adenovirus capsid hexon genotyping was performed by PCR and sequencing using a specified primer set (ADHEX1F/AD2) or a different primer set (AD1/AD2) [10 (link)]. Additionally, astrovirus and sapovirus genotyping was performed by PCR and sequencing techniques using specific capsid primer sets [11 (link)12 (link)]. The PCR products were visualized by electrophoresis on an agarose gel and analyzed by DNA sequencing. The nucleotide sequences were analyzed using ABI Prism BigDye Terminator version 3.1 (Applied Biosystems, Foster City, CA, USA), and the genotypes were confirmed using the NCBI BLAST server of the GenBank database.
Protocol detail
Genotyping of Viral Capsid Genes
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Abi 1
Adenovirus
Agarose
Astrovirus
Capsid
Electrophoresis
Genes
Genotypes
Hexon
Norovirus
Nucleotide sequences
Primer
Prism
Rotavirus g
Sapovirus
Corresponding Organization :
Other organizations : Hallym University
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Variable analysis
independent variables
- Primer sets for rotavirus G (VP7) and P (VP4) genotyping
- Primer sets for norovirus capsid genotyping
- Primer sets for adenovirus capsid hexon genotyping
- Primer sets for astrovirus and sapovirus genotyping
- Rotavirus G (VP7) and P (VP4) genotypes
- Norovirus capsid genotypes
- Adenovirus capsid hexon genotypes
- Astrovirus and sapovirus genotypes
- PCR and sequencing methods for genotyping
- Visualization of PCR products by electrophoresis on an agarose gel
- DNA sequence analysis using ABI Prism BigDye Terminator version 3.1
- Genotype confirmation using the NCBI BLAST server of the GenBank database
- Not explicitly mentioned
- Not explicitly mentioned
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