Optimization of virus detection in cells using massively parallel sequencing

摘要: Massively parallel sequencing (MPS)-based virus detection has potential regulatory applications. We studied the ability of one these approaches, based on degenerate oligonucleotide primer (DOP)-polymerase chain reaction (PCR), to detect viral sequences in cell lines known express genes or particles. DOP-PCR was highly sensitive for small quantities isolated sequences. Detected included nodavirus, bracovirus, and endogenous retroviruses High Five cells, porcine circovirus type 1 retrovirus PK15 human T-cell leukemia MJ papillomavirus 18 HeLa herpesvirus 8 BCBL-1 Epstein–Barr Virus Raji cells. Illumina (for which primers were most efficiently added using PCR) provided greater sensitivity than Roche 454 sequencing. Analyzing nucleic acids extracted both directly from samples capsid-enriched preparations useful information. Although there are limitations methods, results indicate significant promise combination nonspecific PCR MPS identifying contaminants clinical biological samples, including reagents used produce vaccines therapeutic products.