Development of a novel digital RT-PCR method for detection of human sapovirus in different matrices

J Virol Methods. 2018 Apr:254:21-24. doi: 10.1016/j.jviromet.2018.01.005.

Abstract

A new nanofluidic digital RT-PCR method was developed for sapovirus (SaV) using control material obtained according to standards for enteric viruses. Primers employed amplify a fragment of 112 bp of the polymerase capsid junction, allowing the detection of human genogroups I, II and IV. Analytical validation was performed in clinical, shellfish and environmental water samples. This novel protocol rendered great effectiveness and repetitiveness, as well as higher sensitivity than real time RT-PCR assay, with differences in quantification ranging from 0.1 to 2.6 log-units. The method described here can constitute a promising tool for standardizing SaV quantification.

Keywords: Absolute quantification; Clinical samples; Digital RT-PCR; Environmental samples; Human sapovirus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Caliciviridae Infections / diagnosis*
  • Caliciviridae Infections / virology*
  • Gastroenteritis / diagnosis*
  • Gastroenteritis / virology*
  • Genotype
  • Humans
  • RNA, Viral
  • Reproducibility of Results
  • Reverse Transcriptase Polymerase Chain Reaction* / methods
  • Sapovirus / genetics*
  • Sapovirus / isolation & purification
  • Sensitivity and Specificity

Substances

  • RNA, Viral