Next-Generation Sequencing for Pathogen Discovery in Meningoencephalitis (P2.306)
Citation Manager Formats
Make Comment
See Comments

Abstract
Objective: Describe a clinical case in which next-generation sequencing (NGS) rapidly identified a rare but treatable bacterial cause of meningoencephalitis resulting in a favorable patient outcome. Background: Over 100 pathogens cause meningoencephalitis, complicating the development of any comprehensive diagnostic workup. Unbiased NGS is revolutionizing our ability to identify new pathogens, especially novel viruses. However, the utility of NGS in the context of critical illness has not been established. Design/Methods: A 14 year-old boy with severe combined immunodeficiency presented with 4 months of headache and fever progressing to hydrocephalus and coma. An exhaustive diagnostic workup including brain biopsy was unrevealing. NGS was performed on the patient’s cerebrospinal fluid (CSF) and blood. The extracted samples were put through a modified TruSeq protocol (Illumina, San Diego, CA) to generate sequencing libraries. The libraries were sequenced on an Illumina MiSeq instrument using 150/350 base pair paired-end sequencing. Using the SNAP nucleotide aligner, the sequences were analyzed using an in-house computational pipeline for viral, bacterial and fungal sequences. Results: The sequencing run yielded 16,375,474 raw reads. In the untreated CSF, 955 reads aligned to the Leptospiraceae bacterial family, and 483 reads aligned to this family in the pre-DNAsed CSF sample. No reads aligned in the pre-DNAsed serum sample. Further analysis showed the reads aligned to Leptospira santarosai with coverage spanning the entire genome. These results were obtained within 48 hours of obtaining the specimens. The Centers for Disease Control's standard PCR assay was negative, but the 16s ribosomal RNA PCR was positive. The patient received appropriate Leptospira anti-microbials and made a dramatic recovery. He is now at home near his pre-morbid baseline. Conclusions: Leptospirosis is a zoonotic bacterial disease that occurs throughout the world. The disease phenotype is widely variable, and meningitis is common. Diagnostic assays for Leptospira are imperfect and rarely pursued. NGS rapidly detected this rare pathogen leading to a dramatic, real-time impact on this patient’s care.
Disclosure: Dr. Wilson has nothing to disclose. Dr. Samayoa has nothing to disclose. Dr. Naccache has nothing to disclose. Dr. Somasekar has nothing to disclose. Dr. Yu has nothing to disclose. Dr. DeRisi has nothing to disclose. Dr. Chiu has nothing to disclose.
Tuesday, April 29 2014, 7:30 am-11:00 am
- Copyright © 2014 by AAN Enterprises, Inc.
Letters: Rapid online correspondence
REQUIREMENTS
If you are uploading a letter concerning an article:
You must have updated your disclosures within six months: http://submit.neurology.org
Your co-authors must send a completed Publishing Agreement Form to Neurology Staff (not necessary for the lead/corresponding author as the form below will suffice) before you upload your comment.
If you are responding to a comment that was written about an article you originally authored:
You (and co-authors) do not need to fill out forms or check disclosures as author forms are still valid
and apply to letter.
Submission specifications:
- Submissions must be < 200 words with < 5 references. Reference 1 must be the article on which you are commenting.
- Submissions should not have more than 5 authors. (Exception: original author replies can include all original authors of the article)
- Submit only on articles published within 6 months of issue date.
- Do not be redundant. Read any comments already posted on the article prior to submission.
- Submitted comments are subject to editing and editor review prior to posting.
You May Also be Interested in
Dr. Jennifer Majersik and Dr. Carlos Garcia-Esperon
Watch
Related Articles
- No related articles found.


