title: Prospective evaluation of a new automated nucleic acid extraction system using routine clinical respiratory specimens.
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.
authors.affiliation: Department of Virology, Federative Institute of Biology, CH Toulouse, France. mengelle.c@chu-toulouse.fr
authors.forename: C
authors.initials: C
authors.lastname: Mengelle
authors.forename: J-M
authors.initials: JM
authors.lastname: Mansuy
authors.forename: K
authors.initials: K
authors.lastname: Sandres-Sauné
authors.forename: C
authors.initials: C
authors.lastname: Barthe
authors.forename: J
authors.initials: J
authors.lastname: Boineau
authors.forename: J
authors.initials: J
authors.lastname: Izopet
mesh_terms: D001331:Automation / Q000379:methods*; D001992:Bronchoalveolar Lavage Fluid / Q000821:virology; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods; D009305:Nasopharynx / Q000821:virology; D009696:Nucleic Acids / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012141:Respiratory Tract Infections / Q000175:diagnosis* / Q000821:virology; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods*; D014773:Virology / Q000379:methods*; D014777:Virus Diseases / Q000175:diagnosis* / Q000821:virology; D014780:Viruses / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D009696:Nucleic Acids