Improved multiplex-PCR and microarray for herpesvirus detection from CSF

J Clin Virol. 2008 Jun;42(2):172-5. doi: 10.1016/j.jcv.2008.01.003. Epub 2008 Mar 4.

Abstract

Background: A multiplex-PCR and microarray-based method was designed for detection of eight herpesviruses from clinical specimens.

Objectives: To improve the method, especially for detection of herpes simplex (HSV) and varicella-zoster viruses (VZV), and to update and validate the method using positive cerebrospinal fluid (CSF) samples.

Study design: A new primer pair for HSV-PCR and four detection oligonucleotides for HSVs were designed. Two new detection oligonucleotides for VZV and additional oligonucleotides for CMV, EBV, HHV-6 and -7 were designed. The new and previous multiplex-PCR and microarray method were tested in parallel with dilution series of commercial herpesvirus DNAs and 20 CSF specimens positive for HSV-1, HSV-2, or VZV.

Results: New method was more sensitive for detection of HSVs and both two new detection oligonucleotides for VZV functioned well at low levels of viral DNA.

Conclusions: The revised HSV-PCR and new HSV- and VZV-oligonucleotides were found to function well and be more sensitive, thereby increasing reliability of the method.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Central Nervous System Viral Diseases / cerebrospinal fluid
  • Central Nervous System Viral Diseases / diagnosis*
  • Central Nervous System Viral Diseases / virology
  • Cerebrospinal Fluid / virology*
  • DNA, Viral / analysis
  • DNA, Viral / cerebrospinal fluid
  • Herpesviridae / genetics
  • Herpesviridae / isolation & purification*
  • Herpesviridae Infections / cerebrospinal fluid
  • Herpesviridae Infections / diagnosis*
  • Herpesviridae Infections / virology
  • Humans
  • Oligonucleotide Array Sequence Analysis / methods*
  • Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity

Substances

  • DNA, Viral