title: Simultaneous Extraction of Viral and Bacterial Nucleic Acids for Molecular Diagnostic Applications.
abstract: Molecular detection of microbial pathogens in clinical samples requires the application of efficient sample lysis protocols and subsequent extraction and isolation of their nucleic acids. Here, we describe a simple and time-efficient method for simultaneous extraction of genomic DNA from gram-positive and -negative bacteria, as well as RNA from viral agents present in a sample. This method compared well with existing bacterial- and viral-specialized extraction protocols, worked reliably on clinical samples, and was not pathogen specific. This method may be used to extract DNA and RNA concurrently from viral and bacterial pathogens present in a sample and effectively detect coinfections in routine clinical diagnostics.
authors.affiliation: 1 Science Applications International Corporation, San Diego, California, USA; and 2 Department of Clinical Investigation, Tripler Army Medical Center, Honolulu, Hawaii, USA.
authors.forename: Lauren N
authors.initials: LN
authors.lastname: Kajiura
authors.affiliation: 1 Science Applications International Corporation, San Diego, California, USA; and 2 Department of Clinical Investigation, Tripler Army Medical Center, Honolulu, Hawaii, USA.
authors.forename: Scott D
authors.initials: SD
authors.lastname: Stewart
authors.affiliation: 1 Science Applications International Corporation, San Diego, California, USA; and 2 Department of Clinical Investigation, Tripler Army Medical Center, Honolulu, Hawaii, USA.
authors.forename: John
authors.initials: J
authors.lastname: Dresios
authors.affiliation: 1 Science Applications International Corporation, San Diego, California, USA; and 2 Department of Clinical Investigation, Tripler Army Medical Center, Honolulu, Hawaii, USA.
authors.forename: Catherine F T
authors.initials: CF
authors.lastname: Uyehara
mesh_terms: D002853:Chromatography, Liquid; D060085:Coinfection / Q000175:diagnosis / Q000821:virology; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D006193:Haemophilus influenzae / Q000235:genetics; D006801:Humans; D053122:Influenza A Virus, H3N2 Subtype / Q000235:genetics; D025202:Molecular Diagnostic Techniques; D016133:Polymerase Chain Reaction; D012329:RNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D013211:Staphylococcus aureus / Q000235:genetics
chemical_list: D004269:DNA, Bacterial; D004279:DNA, Viral; D012329:RNA, Bacterial; D012367:RNA, Viral
keywords: PCR; RNA/DNA extraction; bacteria; coinfection; mass spectrometry; virus