New LightCycler PCR for rapid and sensitive quantification of parvovirus B19 DNA guides therapeutic decision-making in relapsing infections

J Clin Microbiol. 2001 Dec;39(12):4413-9. doi: 10.1128/jcm.39.12.4413-4419.2001.

Abstract

Detection of parvovirus B19 DNA offers diagnostic advantages over serology, particularly in persistent infections of immunocompromised patients. A rapid, novel method of B19 DNA detection and quantification is introduced. This method, a quantitative PCR assay, is based on real-time glass capillary thermocycling (LightCycler [LC]) and fluorescence resonance energy transfer (FRET). The PCR assay allowed quantification over a dynamic range of over 7 logs and could quantify as little as 250 B19 genome equivalents (geq) per ml as calculated for plasmid DNA (i.e., theoretically >or=5 geq per assay). Interrater agreement analysis demonstrated equivalence of LC-FRET PCR and conventional nested PCR in the diagnosis of an active B19 infection (kappa coefficient = 0.83). The benefit of the new method was demonstrated in an immunocompromised child with a relapsing infection, who required an attenuation of the immunosuppressive therapy in addition to repeated doses of immunoglobulin to eliminate the virus.

Publication types

  • Case Reports
  • Evaluation Study

MeSH terms

  • Child
  • DNA, Viral / blood*
  • Energy Transfer
  • Female
  • Fluorescence
  • Humans
  • Immunocompetence
  • Immunocompromised Host
  • Parvoviridae Infections / prevention & control
  • Parvoviridae Infections / virology*
  • Parvovirus B19, Human / genetics
  • Parvovirus B19, Human / isolation & purification*
  • Parvovirus B19, Human / physiology*
  • Polymerase Chain Reaction* / instrumentation
  • Polymerase Chain Reaction* / methods
  • Recurrence
  • Sensitivity and Specificity
  • Spectrometry, Fluorescence
  • Time Factors

Substances

  • DNA, Viral