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Detection of influenza by real time RT-PCR is not affected by delays in respiratory specimen processing

tetano

Editor, Senior Moderator
J Med Virol. 2016 Apr 18. doi: 10.1002/jmv.24549. [Epub ahead of print]
[h=1]Detection of influenza by real time RT-PCR is not affected by delays in respiratory specimen processing.[/h] Dare R[SUP]1[/SUP], Zhu Y[SUP]2[/SUP], Williams JV[SUP]3[/SUP], Griffin M[SUP]1,[/SUP][SUP]4,[/SUP][SUP]5[/SUP], Talbot HK[SUP]1[/SUP].
[h=3]Author information[/h]

[h=3]Abstract[/h] [h=4]BACKGROUND:[/h] 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.
[h=4]STUDY DESIGN:[/h] 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.
[h=4]RESULTS:[/h] 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.
[h=4]CONCLUSION:[/h] 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. This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.


[h=4]KEYWORDS:[/h] Influenza; Processing; Respiratory; Swabs; real-time RT PCR

PMID: 27089468 [PubMed - as supplied by publisher]
 
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