Comparison of specific and random priming in the reverse transcriptase polymerase chain reaction for genotyping group A rotaviruses

G2P[4] the most prevalent rotavirus genotype in 2007 winter season in an European non-vaccinated population

“…The majority of the mixed infections were with G1 and G2 RV strains, 54.5% of the mixed infections (Table 2). Unusual G/P associations were observed: 1-G1P [4], 2-G2P [8], 1-G4P [9], 1-G9P [4], and 2-G9P [9].…”

“…Reverse transcription was performed as previously described. 8,9 The genotypes were determined in Braga, Portugal, and confirmed in the UK (Virus Reference Department, Health Protection Agency, London).…”

Frequency, clinical characteristics, and genotype distribution of rotavirus gastroenteritis in Greece (2007–2008)

“…Viral RNA was extracted from 10% faecal suspensions in balanced salt solution with the guanidinium isothiocyanate/silica method [Boom et al, 1990]. Complementary DNA was synthesized from the extracted viral RNA using reverse transcriptase enzyme in the presence of random hexamers [Iturriza-Gómara et al, 1999] and was further characterised by semi-nested multiplex polymerase chain reaction (PCR) using specific primers for the VP7 and VP4 genes to identify the G and P types respectively [Gentsch et al, 1992;Gouvea et al, 1990;Iturriza-Gómara et al, 2000a;IturrizaGómara et al, 2004;Simmonds et al, 2008]. Amplification products were analysed by electrophoresis using a 2% agarose gel stained with ethidium bromide in TBE buffer.…”

Hospitalization due to norovirus and genotypes of rotavirus in pediatric patients, state of Espírito Santo

“…Complementary DNA (cDNA) was obtained in a reverse transcription reaction using 20 mU pd(N)6™ (Amersham Bioscience, UK) hexanucleotide random primer (Iturriza-Gomara et al 1999), after denaturation with dimethylsulfoxide for 7 min at 97ºC. The cDNA was then submitted to nucleic acid amplification for NoV detection in all samples and for RV genotypes of those that were positive on PAGE.…”