Detection of Zika virus by SYBR green one-step real-time RT-PCR

J Virol Methods. 2016 Oct:236:93-97. doi: 10.1016/j.jviromet.2016.07.014. Epub 2016 Jul 18.

Abstract

The ongoing Zika virus (ZIKV) outbreak has rapidly spread to new areas of Americas, which were the first transmissions outside its traditional endemic areas in Africa and Asia. Due to the link with newborn defects and neurological disorder, numerous infected cases throughout the world and various mosquito vectors, the virus has been considered to be an international public health emergency. In the present study, we developed a SYBR Green based one-step real-time RT-PCR assay for rapid detection of ZIKV. Our results revealed that the real-time assay is highly specific and sensitive in detection of ZIKV in cell samples. Importantly, the replication of ZIKV at different time points in infected cells could be rapidly monitored by the real-time RT-PCR assay. Specifically, the real-time RT-PCR showed acceptable performance in measurement of infectious ZIKV RNA. This assay could detect ZIKV at a titer as low as 1PFU/mL. The real-time RT-PCR assay could be a useful tool for further virology surveillance and diagnosis of ZIKV.

Keywords: Real-time RT-PCR; SYBR green; Zika virus.

Publication types

  • Evaluation Study

MeSH terms

  • Animals
  • Benzothiazoles
  • Cell Line
  • Diamines
  • Humans
  • Organic Chemicals / metabolism*
  • Quinolines
  • RNA, Viral / analysis*
  • RNA, Viral / genetics
  • Real-Time Polymerase Chain Reaction / methods*
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Staining and Labeling / methods*
  • Zika Virus / genetics
  • Zika Virus / isolation & purification*

Substances

  • Benzothiazoles
  • Diamines
  • Organic Chemicals
  • Quinolines
  • RNA, Viral
  • SYBR Green I