Metagenomics Using Next-Generation Sequencing for Biological Products

Background and Aim: Metagenomics technology enables sequencingof biological material with a flow much higher than before. Thus, thearrival of this technology allows microbiomes of very diverse habitatsto be described. The fetal bovine serum is a potential source of viralcontamination for cell cultures used in the production of biologicalproducts for human or animal use and is therefore routinely subjected to arange of virus-specific tests to ensure an absence of viral contaminations.Methods: Using viral metagenomics we describe here the virome inserum from calves. For this purpose, collected samples are to be classified, homogenized, filtered, and then treated with nucleases prior to nucleicacid extraction. Following random RT-PCR and use of Nextera reagents(Illumina), we will generate doubly tagged DNA from each supernatantfor sequencing. Sequence reads will be assembled de novo by use of theEnsemble program, and computational analysis will be performed usinga designed pipeline.Results: The nearly complete genomes of different virus speciesincluding some from potentially novel RNA and DNA viral families arecharacterized and phylogenetically analyzed, significantly expanding theknown serum-associated virome.Conclusion: This study further characterizes the diversity of viruses in calfserum with the potential to infect fetuses and through fetal bovine serumcontaminate cell cultures.

کلیدواژه ها:

Next-generation sequencing ، Cell culture ، Virus