title: One-step quantitative RT-PCR for the detection of rotavirus in acute gastroenteritis.
abstract: The standard diagnosis of rotavirus gastroenteritis is based on the demonstration of rotavirus antigen in stools using an enzyme immunoassay (EIA). In this study, a one-step quantitative RT-PCR (Q-PCR) was used for sensitive detection of rotavirus in diarrheal stools. The primers and TaqMan probe for the Q-PCR were selected from a highly conserved region of the non-structural protein 3 (NSP3) of rotavirus. After validation, the test was applied to study rotavirus EIA positive (N=25) and EIA negative (N=143) stool specimens from cases of acute gastroenteritis of all degrees of severity in a prospective follow-up cohort of infants from 2 months to 2 years of age. Q-PCR detected all 25 EIA positive rotavirus antigens and seven additional cases that were rotavirus EIA negative, i.e. 28% more rotavirus positive cases than identified by EIA. It is concluded that Q-PCR using primers targeted at NSP3 is a rapid and sensitive method for diagnosing acute rotavirus gastroenteritis.
authors.affiliation: Vaccine Research Center, University of Tampere, Medical School, Vaccine Research Center, Biokatu 10, FIN-33520 Tampere, Finland. shangqin.zeng@uta.fi
authors.forename: S Q
authors.initials: SQ
authors.lastname: Zeng
authors.forename: A
authors.initials: A
authors.lastname: Halkosalo
authors.forename: M
authors.initials: M
authors.lastname: Salminen
authors.forename: E D
authors.initials: ED
authors.lastname: Szakal
authors.forename: L
authors.initials: L
authors.lastname: Puustinen
authors.forename: T
authors.initials: T
authors.lastname: Vesikari