Impact of nucleic acid extraction platforms on hepatitis virus genome detection

Details

• Alternative Title:
  J Virol Methods
• Personal Author:
  Pauly, Matthew D. ; Kamili, Saleem ; Hayden, Tonya M.
• Description:
  Detection and quantification of viral nucleic acids are important for diagnosing current viral infections and monitoring response to antiviral therapy. Automated nucleic acid extraction and purification platforms are routinely used during the first step in these processes in clinical and research laboratories. Here, we compare the extraction efficiencies of four MagNA Pure magnetic bead-based nucleic acid extraction platforms and associated kits using samples positive for nucleic acids from HAV, HBV, HCV, HDV, and HEV. These five hepatitis viruses are diverse in their virion structures and type of nucleic acid that compose their genomes. We found that the most efficient nucleic acid extraction platform and corresponding kit, when averaged across all tested viruses, was the MagNA Pure 96, which yielded twice as much detectable nucleic acid as the other platforms. However, the relative efficiencies of the different platforms varied by virus type, suggesting that an extraction platform that is more efficient for one virus type will not necessarily function better with a different virus type. Our results show that the choice of a nucleic acid extraction platform influences the sensitivity of the methodology and has the potential to generate false-negative results especially in samples with low levels of viral nucleic acids.
• Subjects:
  Article DNA, Viral Genome, Viral Hepatitis, Viral, Human Hepatitis Viruses Humans Nucleic Acids Nucleic Acid Testing Reagent Kits, Diagnostic RNA, Viral Sensitivity And Specificity Viral Hepatitis

  More +
• Source:
  J Virol Methods. 273:113715
• Pubmed ID:
  31419455
• Pubmed Central ID:
  PMC6836433
• Document Type:
  Journal Article
• Funding:
  CC999999/ImCDC/Intramural CDC HHS/United States
• Volume:
  273
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:daf9f5484617d054cbd5f6cf6bcdb8fd7565851a0e76cc5d0091334fbca1baeb
• Download URL:
  https://stacks.cdc.gov/view/cdc/86385/cdc_86385_DS1.pdf
• File Type:
  [PDF - 756.75 KB ]