Validation of MSwab medium long term frozen stability of clinical specimens for the detection of viruses by nucleic acids amplification assays and by culture

No: 1607 Presentation at ESCV 2015: Poster 2 Validation of MSwab medium long term frozen stability of clinical specimens for the detection of viruses by nucleic acids amplification assays and by culture S. Castriciano1, C. Luinstra2, C. Rutherford2, M. Booth2, M. Smieja2,3 1 Copan Italia Spa, Italy 2 St. Joseph’s Healthcare, Hamilton ON, Canada 3 Department of Pathology u0026 Molecular Medicine, McMaster University, Canada Background: MSwab is a molecular medium for the collection, and storage of clinical specimens for the detection of viruses with direct rapid or traditional nucleic acid extraction and amplification assays and culture. The objectives of this studywere to validate the performance of the MSwabTM for: (1) Direct rapid and traditional nucleic acids extraction for the detection of viruses by real time PCR. (2) Virus isolation by shell vial culture. (3) Viruses long term frozen stability inMSwab for direct rapid and traditional nucleic acid extraction and detection by real time PCR and culture. Methods: The flocked swab from Nasopharyngeal (NP) and lesion swab (LS) in UTM (Copan Italia) submitted for virus detection was used for this validation. After the samples were tested by real time PCR, the flocked swabs from positive swab samples were transferred to a 1.6ml tube of MSwab medium. Five positive each for influenza A (FA), influenza B, (FB) Respiratory Syncytial Virus (RSV), Parainfluenza type 1, 2, and 3 (P1, P2, P3), Adenovirus (ADV), human metapneumovirus (hMPV) Herpes simplex virus 1 and 2 (HSV1, HSV2), and Varicella zoster virus (VZV), were used. Each MSwab medium tube with the positive swabs was vortexed and 200 l of MSwab sample were used to inoculate a shell vial culture, another 200 l were added to a microtube and placed in a dry heating block at 980–1000C for 5min, vortexed for 10 s and centrifuged at 14,000 rpm for 2min. A 200 l aliquot was extracted with the easyMagTM (bioMerieux) protocol and eluted in 55 l. Five microliters of nucleic acids extracted with both rapid and traditionalmethodswere testedwith the in-housemultiplex real time PCR and compared to the UTM reference results. Left over MSwab sampleswere storedat -80degreeCand testedafter1year as above. Results: MSwab direct rapid and EasyMag nucleic acid extraction tested with multiplex real time PCR and shell vial culture confirmed all five FA FB, RSV, P1, P2, P3, ADV, HMPV, HSV1, HSV2 and VZV. No toxicity or contamination was observed in culture. Viruses’ viability and nucleic acids were stable in MSwab after 1 year at −80 ◦C. Conclusions: MSwab direct rapid nucleic acid extraction detected all positive tested by real time PCR and culture at collection time and after one year storage at −80 ◦C. MSwab direct rapid extraction improves results turnaround time, save costly extraction reagents and supports culture confirmation. http://dx.doi.org/10.1016/j.jcv.2015.07.231 Abstract No: 1609 Presentation at ESCV 2015: Poster 2 Harnessing an innate immune mechanism to cure viral infections. The mechanism behind viral inhibition by NaCl B. Cai1, S. Griffiths1, J. Wong1, M. Twomey1, R. Chen1, J.G. Haas1, S. Ramalingam1,2 1 Division of Infection and Pathway Medicine, University of Edinburgh, UK 2 Department of Laboratory Medicine, NHS Lothian,