Next-Generation Sequencing for Diagnosis and Tailored Therapy: A Case Report of Astrovirus-Associated Progressive Encephalitis

Frémond, M.-L.; Pérot, Philippe; Muth, Erika; Cros, Guilhem; Dumarest, Marine; Mahlaoui, Nizar; Seilhean, Danielle; Desguerre, Isabelle; Hébert, Charles; Corre-Catelin, Nicole; Neven, Bénédicte; Lecuit, Marc; Blanche, Stéphane; Pïcard, Capucine; Éloit, Marc

doi:10.1093/jpids/piv040

Cited by 122 publications

(132 citation statements)

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“…These data were corroborated in two of the cases by immunohistochemistry and in another case by in situ hybridization, localizing VA1 to neurons and astrocytes (32)(33)(34). Death occurred in four of five patients, demonstrating a high mortality rate in the reported VA1-associated cases of encephalitis (32)(33)(34)(35)(36).…”

supporting

confidence: 58%

“…In these cases, all patients were immunocompromised, including three hematopoietic stem cell transplant recipients and two patients with X-linked agammaglobulinemia (32)(33)(34)(35)(36). VA1 sequences were detected in brain tissue biopsy specimens of all five cases by unbiased next-generation sequencing (32)(33)(34)(35)(36). These data were corroborated in two of the cases by immunohistochemistry and in another case by in situ hybridization, localizing VA1 to neurons and astrocytes (32)(33)(34).…”

mentioning

confidence: 64%

“…VA1 is the most frequently identified genotype of astrovirus in cases of human encephalitis, with a total of five independently published cases to date (32)(33)(34)(35)(36). In these cases, all patients were immunocompromised, including three hematopoietic stem cell transplant recipients and two patients with X-linked agammaglobulinemia (32)(33)(34)(35)(36).…”

mentioning

confidence: 99%

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Propagation of Astrovirus VA1, a Neurotropic Human Astrovirus, in Cell Culture

Janowski

Bauer

Holtz

et al. 2017

J Virol

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Astrovirus VA1/HMO-C (VA1; mamastrovirus 9) is a recently discovered astrovirus genotype that is divergent from the classic human astroviruses (mamastrovirus 1). The gastrointestinal tract is presumed to be the primary site of infection and pathogenicity for astroviruses. However, VA1 has been independently detected in brain tissue of five cases of human encephalitis. Studies of the pathogenicity of VA1 are currently impossible because there are no reported cell culture systems or in vivo models that support VA1 infection. Here, we describe successful propagation of VA1 in multiple human cell lines. The initial inoculum, a filtered clinical stool sample from the index gastroenteritis case cluster that led to the discovery of VA1, was first passaged in Vero cells. Serial blind passage in Caco-2 cells yielded increasing copies of VA1 RNA, and multistep growth curves demonstrated a Ͼ100-fold increase in VA1 RNA 72 h after inoculation. The full-length genomic and subgenomic RNA strands were detected by Northern blotting, and crystalline lattices of viral particles of ϳ26-nm diameter were observed by electron microscopy in infected Caco-2 cells. Unlike other human astrovirus cell culture systems, which require addition of exogenous trypsin for continued propagation, VA1 could be propagated equally well with or without the addition of trypsin. Furthermore, VA1 was sensitive to the type I interferon (IFN-I) response, as VA1 RNA levels were reduced by pretreatment of Caco-2 cells with IFN-␤1a. The ability to propagate VA1 in cell culture will facilitate studies of the neurotropism and neuropathogenesis of VA1.IMPORTANCE Astroviruses are an emerging cause of central nervous system infections in mammals, and astrovirus VA1/HMO-C is the most prevalent astrovirus in cases of human encephalitis. This virus has not been previously propagated, preventing elucidation of the biology of this virus. We describe the first cell culture system for VA1, a key step necessary for the study of its ability to cause disease.KEYWORDS astrovirus, astrovirus VA1, cell culture, electron microscopy, encephalitis, pathogenesis, subgenomic RNA, viral propagation I nitially identified in 1975, members of the single-stranded, positive-sense RNA viral family Astroviridae have been frequently detected in vertebrate stool samples (1, 2). In humans, the first identified astrovirus species was mamastrovirus 1 (classic human astrovirus), and eight serotypes of this virus have been described (2-4). Most humans are exposed to the classic human astroviruses with seroprevalence as high as 94% to specific serotypes (5-9). The classic human astroviruses predominantly cause a selflimited gastrointestinal illness and have been characterized as the third-to fifth-mostcommon viral etiology of diarrhea and gastroenteritis in humans (10)(11)(12)(13)(14).Astroviruses contain three open reading frames (ORFs) and based on conserved genomic elements are hypothesized to share common mechanisms of replication (2). Astroviruses encode a slippery sequence and stem-...

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supporting

confidence: 58%

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confidence: 64%

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Propagation of Astrovirus VA1, a Neurotropic Human Astrovirus, in Cell Culture

Janowski

Bauer

Holtz

et al. 2017

J Virol

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“…24,[46][47][48][49][50][51][52] Total RNA is extracted from a patient's CSF, and complementary DNA (cDNA) is generated by reverse transcription with random hexamer primers. The use of mNGS has gained momentum over the past several years, with several notable case reports and case series showcasing mNGS's ability to capture a broad range of infections with a single assay.…”

Section: Research Csf Metagenomic Sequencingmentioning

confidence: 99%

Clinicopathology conference: 41‐year‐old woman with chronic relapsing meningitis

Beck

Ramachandran

Khan

et al. 2019

Annals of Neurology

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“…More recently, we and others have shown that improvements of the sample preparation and bioIT pipelines allow actionable diagnostics in a short and constant time. [9,10] These studies have shown the medical benefit of WG-NGS as a "last intention" test in patients who tested negative by standard diagnostic, leading to adapted therapy. In all cases, positive results were not due to the discovery of new pathogens, but to the identification of known pathogens not considered by the physicians as possible etiology or for which corresponding diagnostic tests were not available in clinical laboratories.…”

mentioning

confidence: 99%