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Depletion of human DNA in spiked clinical specimens to improve the sensitivity of pathogen detection by next generation sequencing.

Mohammad R Hasan, Arun Rawat, Patrick Tang, Puthen V Jithesh, Eva Thomas, Rusung Tan, Peter Tilley
Next-generation sequencing (NGS) technology has shown promise for the detection of human pathogens from clinical samples. However, one of the major obstacles to the use of NGS in diagnostic microbiology is the low ratio of pathogen DNA to human DNA in most clinical specimens. In this study, we aimed to develop a specimen-processing protocol to remove human DNA and enrich specimens for bacterial and viral DNA for shotgun metagenomic sequencing. Cerebrospinal fluid (CSF) and nasopharyngeal aspirate (NPA) specimens, spiked with control bacterial and viral pathogens, were processed using either a commercially available kit (MolYsis) or various detergents followed by DNase prior to the extraction of DNA...
April 2016: Journal of Clinical Microbiology
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