Full Molecular Typing of Neisseria meningitidis Directly from Clinical Specimens for Outbreak Investigation
Mark Itsko, Adam C. Retchless, Sandeep J. Joseph, Abigail Norris Turner, José A. Bazan, Adodo Yao Sadji, Rasmata Ouedraogo-Traoré, Xin Wang
Abstract
Neisseria meningitidis is a leading cause of bacterial meningitis and sepsis worldwide and an occasional cause of meningococcal urethritis. When isolates are unavailable for surveillance or outbreak investigations, molecular characterization of pathogens needs to be performed directly from clinical specimens, such as cerebrospinal fluid (CSF), blood, or urine. However, genome sequencing of specimens is challenging because of low bacterial and high human DNA abundances. We developed selective whole-genome amplification (SWGA), an isothermal multiple-displacement amplification-based method, to efficiently enrich, sequence, and de novo assemble N. meningitidis DNA from clinical specimens with low bacterial loads.