Actionable Diagnosis of Neuroleptospirosis by Next-Generation Sequencing

Wilson, Michael R.; Naccache, Samia N.; Samayoa, Erik; Biagtan, Mark; Bashir, Hiba; Yu, Guixia; Salamat, Shahriar; Somasekar, Sneha; Federman, Scot; Miller, Steve; Sokolic, Robert A.; Garabedian, Elizabeth; Candotti, Fabio; Buckley, Rebecca H.; Reed, Kurt D.; Meyer, Teresa; Seroogy, Christine M.; Galloway, Renee L.; Henderson, Sheryl L.; Gern, James E.; DeRisi, Joseph L.; Chiu, Charles Y.

doi:10.1056/nejmoa1401268

Cited by 827 publications

(697 citation statements)

References 34 publications

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“…Samples were processed for MDS analysis as previously described. 1 Briefly, total ribonucleic acid (RNA) was extracted from CSF. Samples were randomly amplified to double stranded cDNA using the NuGEN Ovation v.2 kit (NuGEN, San Carlos, CA) for low nucleic acid content samples, and MDS libraries were constructed using the Nextera protocol (Illumina, San Diego, CA) as previously described.…”

Section: Methodsmentioning

confidence: 99%

Diagnosing Balamuthia mandrillarisEncephalitis With Metagenomic Deep Sequencing

Wilson

Shanbhag

Reid

et al. 2015

Annals of Neurology

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121

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Objective Identification of a particular cause of meningoencephalitis can be challenging due to the myriad bacteria, viruses, fungi, and parasites that can produce overlapping clinical phenotypes, frequently delaying diagnosis and therapy. Metagenomic deep sequencing (MDS) approaches to infectious disease diagnostics are known for their ability to identify unusual or novel viruses and thus are well suited for investigating possible etiologies of meningoencephalitis. Methods We present the case of a 74 year-old woman with endophthalmitis followed by meningoencephalitis. MDS of her cerebrospinal fluid (CSF) was performed to identify an infectious agent. Results Sequences aligning to Balamuthia mandrillaris ribosomal RNA genes were identified in the CSF via MDS. Polymerase chain reaction (PCR) subsequently confirmed the presence of B. mandrillaris in CSF, brain tissue, and vitreous fluid from the patient’s infected eye. B. mandrillaris serology and immunohistochemistry for free-living amoebas on the brain biopsy tissue were positive. Interpretation The diagnosis was made using MDS after the patient had been hospitalized for several weeks and subjected to costly and invasive testing. MDS a powerful diagnostic tool with the potential for rapid and unbiased pathogen identification leading to early therapeutic targeting.

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Section: Methodsmentioning

confidence: 99%

Diagnosing Balamuthia mandrillarisEncephalitis With Metagenomic Deep Sequencing

Wilson

Shanbhag

Reid

et al. 2015

Annals of Neurology

Self Cite

121

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“…The diagnosis of severe leptospirosis through clinical assays such as traditional culture, serologic tests and pathogen-specific PCR assays is difficult due to unsatisfactory sensitivity and specificity [6]. The development of nextgeneration sequencing technique has revolutionized our ability to discover emerging pathogens, especially new viruses [7].…”

Section: Archives Of Clinical Microbiology Issn 1989-8436mentioning

confidence: 99%

A Severe Leptospira interrogans Serovar Copenhageni Infection Diagnosed by Next- Generation Sequencing and Treated with Corticosteroids

Zeng¹,

Liu²,

Yuhai³

et al. 2017

Arch Clin Microbiol

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“…Indeed, there are numerous instances in the literature using metagenomic unbiased sequencing to adjudicate clinical diagnosis. One notable case report from Dr. Charles Chiu's laboratory emphasizes the utility of pathogen agnostic NGS to identify a previously undiagnosed case of leptospirosis, providing a perfect example of NGS as a diagnostic last resort (1 ). While efficacious in this instance, unbiased NGS requires a substantial amount of sequence depth to separate low prevalence pathogen from the overwhelming contribution of host nucleic acid.…”

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confidence: 99%