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Eurosurveillance: Does the length of specimen storage affect influenza testing results by real-time reverse transcription-polymerase chain reaction? An analysis of influenza surveillance specimens, 2008 to 2010

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  • Eurosurveillance: Does the length of specimen storage affect influenza testing results by real-time reverse transcription-polymerase chain reaction? An analysis of influenza surveillance specimens, 2008 to 2010

    Eurosurveillance, Volume 19, Issue 36, 11 September 2014
    Research articles
    Does the length of specimen storage affect influenza testing results by real-time reverse transcription-polymerase chain reaction? An analysis of influenza surveillance specimens, 2008 to 2010
    D L Caselton ()1, G Arunga1, G Emukule1, P Muthoka2, L Mayieka1, A Kosgey1, R Ochola1, L W Waiboci1, D R Feikin3, J A Mott1, R F Breiman3, M A Katz1

    Influenza Program, Global Disease Detection Division, Centers for Disease Control and Prevention, Kenya
    Kenya Ministry of Public Health and Sanitation, Kenya
    International Emerging Infections Program, Global Disease Detection Division, Centers for Disease Control and Prevention, Kenya

    Citation style for this article: Caselton DL, Arunga G, Emukule G, Muthoka P, Mayieka L, Kosgey A, Ochola R, Waiboci LW, Feikin DR, Mott JA, Breiman RF, Katz MA. Does the length of specimen storage affect influenza testing results by real-time reverse transcription-polymerase chain reaction? An analysis of influenza surveillance specimens, 2008 to 2010. Euro Surveill. 2014;19(36):pii=20893. Available online: http://www.eurosurveillance.org/View...rticleId=20893
    Date of submission: 30 May 2013

    In some influenza surveillance systems, timely transport to laboratories for reverse transcription-polymerase chain reaction (RT-PCR) testing is challenging. Guidelines suggest that samples can be stored at 4?C for up to 96 hours but the effect of longer storage times has not been systematically evaluated. We collected nasopharyngeal and oropharyngeal specimens from patients in Kenya and stored them in viral transport medium at 2 to 8?C before testing for influenza A and B using real-time RT-PCR. From April 2008 to November 2010, we collected 7,833 samples; 940 (12%) were positive for influenza. In multivariable analysis, specimens stored for six days were less likely to be influenza-positive compared to specimens stored between zero and one day (adjusted odds ratio (aOR): 0.49, 95% confidence interval (CI): 0.27?0.93). There was no statistically significant difference in influenza positivity of specimens stored for five days compared to zero to one day. There was no statistically significant relationship between days in refrigeration and cycle threshold (Ct) values for positive samples (p=0.31). We found that samples could remain in storage for at least five days without affecting the proportion-positive of samples, potentially increasing the feasibility of including influenza surveillance sites in remote areas.

    In some influenza surveillance systems, timely transport to laboratories for reverse transcription-polymerase chain reaction (RT-PCR) testing is challenging. Guidelines suggest that samples can be stored at 4°C for up to 96 hours but the effect of longer storage times has not been systematically evaluated. We collected nasopharyngeal and oropharyngeal specimens from patients in Kenya and stored them in viral transport medium at 2 to 8°C before testing for influenza A and B using real-time RT-PCR. From April 2008 to November 2010, we collected 7,833 samples; 940 (12%) were positive for influenza. In multivariable analysis, specimens stored for six days were less likely to be influenza-positive compared to specimens stored between zero and one day (adjusted odds ratio (aOR): 0.49, 95% confidence interval (CI): 0.27-0.93). There was no statistically significant difference in influenza positivity of specimens stored for five days compared to zero to one day. There was no statistically significant relationship between days in refrigeration and cycle threshold (Ct) values for positive samples (p=0.31). We found that samples could remain in storage for at least five days without affecting the proportion-positive of samples, potentially increasing the feasibility of including influenza surveillance sites in remote areas. .
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