Detection of Flaviviruses by Reverse Transcriptase-Polymerase Chain Reaction With the Universal Primer Set

Microbiol Immunol. 1997;41(3):209-13. doi: 10.1111/j.1348-0421.1997.tb01192.x.

Abstract

Using a universal primer set designed to match the sequence of the NS1 gene of flaviviruses, the virus RNA of dengue (DEN), Japanese encephalitis (JEV), powassan and langat of Flaviviridae were successfully amplified by polymerase chain reaction (PCR) via cDNA; and with different internal primers, the serotypes of the dengue viruses were identified. Of the 78 clinically diagnosed dengue fever patients, 18 patients were positive for DEN 1, 48 patients for DEN 2 and 8 patients concurrently infected with DEN 4. Of the 52 patients admitted with Japanese encephalitis (JE), 45 were determined to be JEV infections. By nested PCR, we completed the identification of flaviviruses within 2 days. The results show that seven primers have a potential value for rapid clinical diagnosis of flavivirus infections.

MeSH terms

  • China / epidemiology
  • DNA Primers*
  • Dengue / blood
  • Dengue / diagnosis
  • Dengue / epidemiology
  • Dengue / genetics
  • Dengue Virus / classification
  • Dengue Virus / genetics
  • Disease Outbreaks
  • Encephalitis, Japanese / blood
  • Encephalitis, Japanese / diagnosis
  • Encephalitis, Japanese / epidemiology
  • Encephalitis, Japanese / genetics
  • Encephalitis, Tick-Borne / blood
  • Encephalitis, Tick-Borne / diagnosis
  • Encephalitis, Tick-Borne / epidemiology
  • Encephalitis, Tick-Borne / genetics
  • Flavivirus Infections / blood
  • Flavivirus Infections / diagnosis*
  • Flavivirus Infections / epidemiology
  • Flavivirus Infections / genetics
  • Humans
  • Polymerase Chain Reaction / methods*
  • RNA, Viral / isolation & purification
  • Serotyping
  • Viral Nonstructural Proteins / genetics

Substances

  • DNA Primers
  • NS1 protein, Flavivirus
  • RNA, Viral
  • Viral Nonstructural Proteins