Quality control implementation for universal characterization of DNA and RNA viruses in clinical respiratory samples using single metagenomic next-generation sequencing workflow

A. Bal; M. Pichon; C. Picard; JS. Casalegno; M. Valette; I. Schuffenecker; L. Billard; S. Vallet; G. Vilchez; V. Cheynet; G. Oriol; S. Assant; Y. Gillet; B. Lina; K. Brengel-Pesce; F. Morfin; L. Josset

The objective of this study was to implement QCs in a single metagenomic protocol and to 99 evaluate it for the detection of a broad panel of DNA After normalization, a pool of libraries (V/V) was made and quantified using universal KAPA 132 library quantification kit (Kapa Biosystems, Wilmington, MA, USA); 1% PhiX genome was 133 added to the quantified library before sequencing with Illumina NextSeq 500 â„¢ platform 134 ( Fig. 1 ). In addition, it should be noticed that our wet-lab process was designed to prevent 135 contaminations as much as possible: reagents were stored and prepared in a DNA-free room; 136 patient samples were opened in a laminar flow hood in a pre-PCR room; after the 137 amplification step, tubes were handled and stored in a post-PCR room. 138