Abstract: Virus discovery combining sequence unbiased amplification with next generation sequencing is now state-of-the-art. We have previously determined that the performance of the unbiased amplification technique which is operational at our institute, VIDISCA-454, is efficient when respiratory samples are used as input. The performance of the assay is, however, not known for other clinical materials like blood or stool samples. Here, we investigated the sensitivity of VIDISCA-454 with feces-suspensions and serum samples that are positive and that have been quantified for norovirus and human immunodeficiency virus type 1, respectively. The performance of VIDISCA-454 in serum samples was equal to its performance in respiratory material, with an estimated lower threshold of 1,000 viral genome copies. The estimated threshold in feces-suspension is around 200,000 viral genome copies. The decreased sensitivity in feces suspension is mainly due to sequences that share no recognizable identity with known sequences. Most likely these sequences originate from bacteria and phages which are not completely sequenced.
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de Vries, M.; Oude Munnink, B.B.; Deijs, M.; Canuti, M.; Koekkoek, S.M.; Molenkamp, R.; Bakker, M.; Jurriaans, S.; van Schaik, B.D.C.; Luyf, A.C.; Olabarriaga, S.D.; van Kampen, A.H.C.; van der Hoek, L. Performance of VIDISCA-454 in Feces-Suspensions and Serum. Viruses 2012, 4, 1328-1334.
de Vries M, Oude Munnink BB, Deijs M, Canuti M, Koekkoek SM, Molenkamp R, Bakker M, Jurriaans S, van Schaik BDC, Luyf AC, Olabarriaga SD, van Kampen AHC, van der Hoek L. Performance of VIDISCA-454 in Feces-Suspensions and Serum. Viruses. 2012; 4(8):1328-1334.
de Vries, Michel; Oude Munnink, Bas B.; Deijs, Martin; Canuti, Marta; Koekkoek, Sylvie M.; Molenkamp, Richard; Bakker, Margreet; Jurriaans, Suzanne; van Schaik, Barbera D. C.; Luyf, Angela C.; Olabarriaga, Silvia D.; van Kampen, Antoine H. C.; van der Hoek, Lia. 2012. "Performance of VIDISCA-454 in Feces-Suspensions and Serum." Viruses 4, no. 8: 1328-1334.