Sequence-independent single-primer-amplification (SISPA) as a screening technique for detecting unexpected RNA viral adventitious agents in cell cultures

The sequence-independent, single-primer amplification (SISPA) enables the random of nucleic acids, allowing detection and genome sequencing different viral agents. This feature SISPA method provides evidence for application it in monitoring presence adventitious RNA viruses cell cultures. We evaluated a challenge virus representing agent Besides, by optimizing our laboratory, we found false-positive results on negative control lanes electrophoresis gels. To investigate sources contamination, were cloned into Escherichia coli cells, sequenced, phylogenetically analyzed. data revealed that can be used as an adjunct to confirm absence unexpected phylogenetic analysis contaminant sequences showed caused acid commercial cDNA synthesis kit reagents, probably tracing back expression plasmids host ribosomal sequences, production enzymes. Therefore, laboratories using methods must constantly aware potentials such contaminations, yielding background noise final NGS reads.