Prospective evaluation of a new automated nucleic acid extraction system using routine clinical respiratory specimens

J Med Virol. 2012 Jun;84(6):906-11. doi: 10.1002/jmv.23281.

Abstract

The aim of the study was to evaluate the MagNA Pure 96™ nucleic acid extraction system using clinical respiratory specimens for identifying viruses by qualitative real-time PCR assays. Three extraction methods were tested, that is, the MagNA Pure LC™, the COBAS Ampliprep™, and the MagNA Pure 96™ with 10-fold dilutions of an influenza A(H1N1)pdm09 sample. Two hundred thirty-nine respiratory specimens, 35 throat swabs, 164 nasopharyngeal specimens, and 40 broncho-alveolar fluids, were extracted with the MagNA Pure 96™ and the COBAS Ampliprep™ instruments. Forty COBAS Ampliprep™ positive samples were also tested. Real-time PCRs were used to identify influenza A and influenza A(H1N1)pdm09, rhinovirus, enterovirus, adenovirus, varicella zoster virus, cytomegalovirus, and herpes simplex virus. Similar results were obtained on RNA extracted from dilutions of influenza A(H1N1)pdm09 with the three systems: the MagNA Pure LC™, the COBAS Ampliprep™, and the MagNA Pure 96™. Data from clinical respiratory specimens extracted with the MagNA Pure 96™ and COBAS Ampliprep™ instruments were in 98.5% in agreement (P < 0.0001) for influenza A and influenza A(H1N1)pdm09. Data for rhinovirus were in 97.3% agreement (P < 0.0001) and in 96.8% agreement for enterovirus. They were in 100% agreement for adenovirus. Data for cytomegalovirus and HSV1-2 were in 95.2% agreement (P < 0.0001). The MagNA Pure 96™ instrument is easy-to-use, reliable, and has a high throughput for extracting total nucleic acid from respiratory specimens. These extracts are suitable for molecular diagnosis with any type of real-time PCR assay.

Publication types

  • Comparative Study
  • Evaluation Study

MeSH terms

  • Automation / methods*
  • Bronchoalveolar Lavage Fluid / virology
  • Humans
  • Molecular Diagnostic Techniques / methods
  • Nasopharynx / virology
  • Nucleic Acids / isolation & purification*
  • Real-Time Polymerase Chain Reaction / methods
  • Respiratory Tract Infections / diagnosis*
  • Respiratory Tract Infections / virology
  • Sensitivity and Specificity
  • Specimen Handling / methods*
  • Virology / methods*
  • Virus Diseases / diagnosis*
  • Virus Diseases / virology
  • Viruses / genetics*
  • Viruses / isolation & purification

Substances

  • Nucleic Acids