title: Evaluation of a multiplex real time reverse transcription PCR assay for the detection and quantitation of the most common human rotavirus genotypes.
abstract: Group A rotaviruses (RVs) are important pathogens that cause acute, dehydrating gastroenteritis in infants and young children. In this study, a multiplex real-time polymerase chain reaction protocol using primers and TaqMan(®) probes specific for viral VP4 and VP7 genes was evaluated. This assay offers simultaneous genotyping and quantification of the most common RV genotypes G1P[8], G2P[4], G3P[8], G4P[8], and G9P[8]. It was compared to the molecular typing results provided by conventional PCR. A total of 92 archived stool specimens obtained from children younger than 5 years old with the diagnosis of acute gastroenteritis were examined. Real-time PCR assay detected rotavirus strains among the most common genotype combinations G4P[8] (70.7%), G1P[8] (10.9%), G2P[4] (5.4%), G9P[8] (2.2%). This new assay described has an acceptable sensitivity (low limit 6.3×10(2)copies/g of stool).
authors.affiliation: Department of Cytopathology, Attikon University Hospital, Athens University Medical School, Athens, Greece. ckottaridi@gmail.com
authors.forename: Christine
authors.initials: C
authors.lastname: Kottaridi
authors.forename: Aris T
authors.initials: AT
authors.lastname: Spathis
authors.forename: Chara Kleio
authors.initials: CK
authors.lastname: Ntova
authors.forename: Vassiliki
authors.initials: V
authors.lastname: Papaevangelou
authors.forename: Petros
authors.initials: P
authors.lastname: Karakitsos
mesh_terms: D000956:Antigens, Viral / Q000235:genetics; D036022:Capsid Proteins / Q000235:genetics; D002675:Child, Preschool; D005759:Gastroenteritis / Q000175:diagnosis* / Q000453:epidemiology / Q000821:virology; D005838:Genotype; D006115:Greece / Q000453:epidemiology; D006801:Humans; D015995:Prevalence; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012401:Rotavirus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D012400:Rotavirus Infections / Q000175:diagnosis* / Q000453:epidemiology / Q000821:virology; D012680:Sensitivity and Specificity
chemical_list: D000956:Antigens, Viral; D036022:Capsid Proteins; D012367:RNA, Viral; C060339:VP4 protein, Rotavirus; C050779:VP7 protein, Rotavirus