Each state and territory independently evaluated which vaccine to implement. Victoria, Queensland, Western Australia and South Australia currently use RotaTeq™, New South Wales, the Northern Territory, Tasmania and the Australian Capital Territory use Rotarix™ [15]. Compound Library Prior to vaccine introduction in Australia, 115,000 GP consults, 22,000 emergency department presentations and 10,000 hospitalisations in children under five years of age could be attributed to rotavirus infection annually [16]. In this study we report the characterisation and molecular analysis
of a G9P[8] strain responsible for a large outbreak of rotavirus gastroenteritis in the Northern Territory of Australia in 2007, five months after the commencement of the Rotarix™ vaccination program. A total of 107 stool samples were collected from paediatric patients hospitalised with severe gastroenteritis
during a rotavirus outbreak in the Alice Springs Osimertinib region of the Northern Territory between the 12th of March and the 11th of July 2007. Patient information including date of birth, date of sample collection, sex and rotavirus immunisation status was obtained. Samples were stored frozen and forwarded to the Australian Rotavirus Reference Centre (ARRC) in Melbourne. Ninety-nine samples had adequate sample for analysis and were characterised using a combination of serotyping EIA and hemi-nested multiplex RT-PCR. Seventy-eight samples were found to be rotavirus positive and typed as G9P[8] and were analysed further in this study [25]. Rotavirus dsRNA was extracted from clarified
20% faecal suspensions using a RNA extraction Kit (QIAamp® Viral RNA mini Kit (spin protocol), Qiagen, Inc., Hilden, Germany) in accordance with the manufacturer’s instructions for use in RT-PCR. Rotavirus dsRNA was extracted from 20% faecal suspensions using phenol-chloroform extraction and purified using hydroxyapatite as previously described for use in Polyacrylamide Gel Electrophoresis (PAGE) [17]. The dsRNA genome segments were separated on 10% (w/v) polyacrylamide gel and the genome migration patterns (electropherotypes) were tuclazepam visualised by silver staining according to the established protocol [18] and [19]. Of the 78 rotavirus positive samples collected during the outbreak, 14 were selected for further analysis including five from vaccinated patients. Samples were evenly selected during the outbreak period. Portions of gene segment 4 (VP4), 9 (VP7) and 10 (NSP4) were reverse transcribed and amplified by PCR using the Superscript III One Step RT-PCR with Platinum Taq DNA Polymerase (Invitrogen, Carlsbad, CA, USA). RNA was denatured and reverse transcribed at 45 °C for 30 min followed by PCR activation at 95 °C for 15 min.
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