Evaluation of a multiplex real time reverse transcription PCR assay for the detection and quantitation of the most common human rotavirus genotypes

J Virol Methods. 2012 Mar;180(1-2):49-53. doi: 10.1016/j.jviromet.2011.12.009. Epub 2012 Jan 11.

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).

Publication types

  • Evaluation Study

MeSH terms

  • Antigens, Viral / genetics
  • Capsid Proteins / genetics
  • Child, Preschool
  • Gastroenteritis / diagnosis*
  • Gastroenteritis / epidemiology
  • Gastroenteritis / virology
  • Genotype
  • Greece / epidemiology
  • Humans
  • Prevalence
  • RNA, Viral / analysis
  • RNA, Viral / isolation & purification
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Rotavirus / classification
  • Rotavirus / genetics*
  • Rotavirus / isolation & purification*
  • Rotavirus Infections / diagnosis*
  • Rotavirus Infections / epidemiology
  • Rotavirus Infections / virology
  • Sensitivity and Specificity

Substances

  • Antigens, Viral
  • Capsid Proteins
  • RNA, Viral
  • VP4 protein, Rotavirus
  • VP7 protein, Rotavirus