title: Nucleotide mismatches between the VP7 gene and the primer are associated with genotyping failure of a specific lineage from G1 rotavirus strains.
abstract: In recent years it was reported that the accumulation of point mutations in VP4 and VP7 genes of rotavirus strains was the main cause of the failure of the G or P-typing. Failures in the correct genotyping of G1, G2, G8, G9 and G10 rotavirus strains were reported in the most commonly used reverse transcription (RT)-PCR strategies. Collecting VP7 gene sequences of G1 rotavirus strains from databases we found that 74 (61.2 %) out of 121 G1 strains from lineage I showed the four specific mismatches at the 5' end of the 9T1-1 primer, previously associated with the failure of G1-typing. Thus, a great percentage of the G1 strains from lineage I worldwide reported could not have been typed if the Das's RT-PCR strategy were used. This analysis shows that the failure on the detection of the G1 strains could be due to the diversification of rotavirus strains in phylogenetic lineages. Therefore, the use of different RT-PCR strategies with different primer binding locations on the VP7 gene or new typing methodologies -like microarrays procedures- could be a better option to avoid the failure of the G-typing of rotavirus strains detected during surveillance programs.
authors.affiliation: Departamento de Biología Molecular, Instituto de Investigaciones en Ciencias de la Salud, Universidad Nacional de Asunción, Río de la Plata y Lagerenza, Asunción (2511), Paraguay. gabriel_parra@hotmail.com
authors.forename: Gabriel I
authors.initials: GI
authors.lastname: Parra
authors.forename: Emilio E
authors.initials: EE
authors.lastname: Espinola
mesh_terms: D000956:Antigens, Viral / Q000235:genetics*; D020137:Base Pair Mismatch; D036022:Capsid Proteins / Q000235:genetics*; D017931:DNA Primers; D005838:Genotype; D006801:Humans; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012401:Rotavirus / Q000145:classification* / Q000235:genetics*
chemical_list: D000956:Antigens, Viral; D036022:Capsid Proteins; D017931:DNA Primers; C050779:VP7 protein, Rotavirus