Differentiation of RotaTeq® vaccine strains from wild-type strains using NSP3 gene in reverse transcription polymerase chain reaction assay

J Virol Methods. 2016 Nov:237:72-78. doi: 10.1016/j.jviromet.2016.08.022. Epub 2016 Aug 30.

Abstract

RotaTeq® is a live attenuated human-bovine reassortant vaccine against rotaviruses that is used worldwide. However, shedding of the virus used in RotaTeq® has been detected in the feces of children following vaccination by the oral route, possibly affecting community immunity. Therefore, a simple and efficient method to discriminate between virulent and RotaTeq® vaccine strains is required. In this study, a novel one-step multiplex reverse-transcription polymerase chain reaction (RT-PCR) assay targeting the NSP3 gene was developed to detect RotaTeq® vaccine strains in fecal samples. RotaTeq® vaccine viruses were successfully distinguished from known wild-type rotavirus genotypes. In addition, the developed assay was able to detect rotaviruses in clinical stool samples obtained from South Korea during the 2011-2013 rotavirus seasons. Of the 1106 stool specimens from children with acute gastroenteritis that were screened, 286 rotaviruses were genotyped. RotaTeq® vaccine strains were identified in 39 samples (13.6%). The novel RT-PCR assay that was developed could be used to detect and discriminate between RotaTeq® vaccine strains that are shed in fecal matter, and to estimate the quantification of virus that has been shed after vaccination.

Keywords: Genotypes; RotaTeq; Rotavirus; Shedding.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle
  • Feces / virology
  • Female
  • Gastroenteritis / virology*
  • Genotype
  • Humans
  • Infant
  • Male
  • Republic of Korea
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Rotavirus / genetics*
  • Rotavirus / isolation & purification*
  • Rotavirus / pathogenicity
  • Rotavirus / physiology
  • Rotavirus Infections / diagnosis
  • Rotavirus Infections / virology*
  • Rotavirus Vaccines / genetics*
  • Vaccines, Attenuated / genetics
  • Viral Nonstructural Proteins / genetics*
  • Virus Shedding

Substances

  • NSP3 protein, Rotavirus
  • RotaTeq
  • Rotavirus Vaccines
  • Vaccines, Attenuated
  • Viral Nonstructural Proteins