The microbial detection array combined with random Phi29-amplification used as a diagnostic tool for virus detection in clinical samples

PLoS One. 2011;6(8):e22631. doi: 10.1371/journal.pone.0022631. Epub 2011 Aug 10.

Abstract

A common technique used for sensitive and specific diagnostic virus detection in clinical samples is PCR that can identify one or several viruses in one assay. However, a diagnostic microarray containing probes for all human pathogens could replace hundreds of individual PCR-reactions and remove the need for a clear clinical hypothesis regarding a suspected pathogen. We have established such a diagnostic platform for random amplification and subsequent microarray identification of viral pathogens in clinical samples. We show that Phi29 polymerase-amplification of a diverse set of clinical samples generates enough viral material for successful identification by the Microbial Detection Array, demonstrating the potential of the microarray technique for broad-spectrum pathogen detection. We conclude that this method detects both DNA and RNA virus, present in the same sample, as well as differentiates between different virus subtypes. We propose this assay for diagnostic analysis of viruses in clinical samples.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacteriophages / enzymology*
  • DNA / genetics
  • DNA Viruses / genetics
  • DNA Viruses / isolation & purification*
  • DNA-Directed DNA Polymerase / metabolism
  • Diagnostic Techniques and Procedures*
  • Genome, Human / genetics
  • Humans
  • Oligonucleotide Array Sequence Analysis / methods*
  • Polymerase Chain Reaction / methods*
  • RNA Viruses / genetics
  • RNA Viruses / isolation & purification*
  • Transcriptome

Substances

  • DNA
  • DNA-Directed DNA Polymerase