Detection of Influenza by Real Time RT-PCR is Not Affected by Delays in Specimen Processing of Respiratory Samples

Details

• Alternative Title:
  J Med Virol
• Personal Author:
  Dare, Ryan ; Zhu, Yuwei ; Williams, John V. ; Griffin, Marie ; Talbot, H. Keipp
• Description:
  Background
  
  Sample processing for real-time reverse transcriptase polymerase chain reaction (rRT-PCR) based diagnostic assays requires stabilizing sample ribonucleic acid (RNA) in lysis buffer prior to testing. The stability of viral RNA prior to processing is difficult to assure. It is unknown if clinical sample integrity is compromised by delays in processing, as may occur due to weekends and holidays. We sought to examine the integrity of respiratory specimens with variable processing times.
  
  Study Design
  
  Upper respiratory specimens were collected during 3 influenza seasons 2009-2012 and tested for influenza virus and internal control human RNase P (RNP) RNA by rRT-PCR. Time to processing was measured in hours from specimen collection to placement in lysis buffer.
  
  Results
  
  635 (11.4%) of 5583 samples were influenza positive. Mean and median times to processing were 11.5 hours and 6.0 hours respectively (min 0.1h, max 105.2h). There were no significant associations between time to processing and presence of RNP (OR=1.0, p=0.740), or detection of influenza (OR=1.0, p=0.060). Longer duration of illness was associated with a lower likelihood of influenza detection (OR=0.92, p<0.001) and with increased influenza A cycle threshold (Ct) values (p<0.001), while older age was associated with increased influenza B Ct values (p=0.001), indicating the presence of less amplifiable RNA.
  
  Conclusion
  
  Delays in time to processing of upper respiratory specimens up to 105 hours were not associated with decreased detection of amplifiable RNA, suggesting specimen integrity is not compromised by such delays.
• Subjects:
  • Age Factors
  • Female
  • Humans
  • Influenza A virus
  • Influenza B virus
  • Influenza, Human
  • Male
  • Middle Aged
  • RNA Stability
  • RNA, Viral
  More +
• Source:
  J Med Virol. 88(11):1891-1895.
• Pubmed ID:
  27089468
• Pubmed Central ID:
  PMC5810138
• Document Type:
  Journal Article
• Funding:
  R01 AG043419/AG/NIA NIH HHS/United States ; UL1 RR024975/RR/NCRR NIH HHS/United States ; U18 IP000184/IP/NCIRD CDC HHS/United States
• Volume:
  88
• Issue:
  11
• Download URL:
  https://stacks.cdc.gov/view/cdc/51689/cdc_51689_DS1.pdf
• File Type:
  [PDF - 154.07 KB]
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:60d5a8de0ef881827f83c210860a726f0cd0fb4bb3504d57dfcef5b1292c8a7f