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Viruses 2014, 6(5), 1876-1896; doi:10.3390/v6051876
Article

Evaluation of the Broad-Range PCR-Electrospray Ionization Mass Spectrometry (PCR/ESI-MS) System and Virus Microarrays for Virus Detection

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Received: 18 December 2013; in revised form: 4 March 2014 / Accepted: 15 April 2014 / Published: 25 April 2014
(This article belongs to the Section Animal Viruses)
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Abstract: Advanced nucleic acid-based technologies are powerful research tools for novel virus discovery but need to be standardized for broader applications such as virus detection in biological products and clinical samples. We have used well-characterized retrovirus stocks to evaluate the limit of detection (LOD) for broad-range PCR with electrospray ionization mass spectrometry (PCR/ESI-MS or PLEX-ID), RT-PCR assays, and virus microarrays. The results indicated that in the absence of background cellular nucleic acids, PLEX-ID and RT-PCR had a similar LOD for xenotropic murine retrovirus-related virus (XMRV; 3.12 particles per µL) whereas sensitivity of virus detection was 10-fold greater using virus microarrays. When virus was spiked into a background of cellular nucleic acids, the LOD using PLEX-ID remained the same, whereas virus detection by RT-PCR was 10-fold less sensitive, and no virus could be detected by microarrays. Expected endogenous retrovirus (ERV) sequences were detected in cell lines tested and known species-specific viral sequences were detected in bovine serum and porcine trypsin. A follow-up strategy was developed using PCR amplification, nucleotide sequencing, and bioinformatics to demonstrate that an RD114-like retrovirus sequence that was detected by PLEX-ID in canine cell lines (Madin-Darby canine kidney (MDCK) and Cf2Th canine thymus) was due to defective, endogenous gammaretrovirus-related sequences.
Keywords: broad-range PCR amplification; electrospray ionization-mass spectrometry; PLEX-ID; virus microarray; RT-PCR; endogenous retroviruses broad-range PCR amplification; electrospray ionization-mass spectrometry; PLEX-ID; virus microarray; RT-PCR; endogenous retroviruses
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Taliaferro, L.P.; Galvin, T.A.; Ma, H.; Shaheduzzaman, S.; Williams, D.K.; Glasner, D.R.; Khan, A.S. Evaluation of the Broad-Range PCR-Electrospray Ionization Mass Spectrometry (PCR/ESI-MS) System and Virus Microarrays for Virus Detection. Viruses 2014, 6, 1876-1896.

AMA Style

Taliaferro LP, Galvin TA, Ma H, Shaheduzzaman S, Williams DK, Glasner DR, Khan AS. Evaluation of the Broad-Range PCR-Electrospray Ionization Mass Spectrometry (PCR/ESI-MS) System and Virus Microarrays for Virus Detection. Viruses. 2014; 6(5):1876-1896.

Chicago/Turabian Style

Taliaferro, Lanyn P.; Galvin, Teresa A.; Ma, Hailun; Shaheduzzaman, Syed; Williams, Dhanya K.; Glasner, Dustin R.; Khan, Arifa S. 2014. "Evaluation of the Broad-Range PCR-Electrospray Ionization Mass Spectrometry (PCR/ESI-MS) System and Virus Microarrays for Virus Detection." Viruses 6, no. 5: 1876-1896.



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