scholarly journals Next-generation sequencing of cerebrospinal fluid for diagnosis of atypical herpes simplex encephalitis

2021 ◽  
Vol 49 (10) ◽  
pp. 030006052110496
Author(s):  
Zhilei Kang ◽  
Xin Jin ◽  
Na Wei ◽  
Ye Ji ◽  
Jingzhe Han
Keyword(s):  
Cerebrospinal Fluid ◽  
Next Generation Sequencing ◽  
Herpes Simplex ◽  
Herpes Simplex Encephalitis ◽  
Clinical Manifestations ◽  
Imaging Features ◽  
Next Generation ◽  
Elevated Protein ◽  
Simplex Virus ◽  
Generation Sequencing

Objectives Herpes simplex encephalitis (HSE) is one of the most common causes of severe viral encephalitis. The characteristic manifestations of HSE include cerebrospinal fluid with mild cytopenia, dominated by lymphocytes, elevated protein, and normal blood glucose values (3.9–6.1 mmol/L). Although it is not difficult to diagnose classical HSE, diagnosing clinically atypical cases is more difficult. Methods We reviewed the results of next-generation sequencing (NGS) of CSF in a series of patients diagnosed with atypical HSE. Results Four patients lacking classical clinical manifestations of HSE, including no fever, headache, or other typical neurological deficit symptoms, 1–2 × 106 cells/L CSF leucocyte count, and no typical imaging features, were diagnosed with atypical HSE by NGS of CSF. The NGS reads corresponding to herpes simplex virus type 1 ranged from 2 to 13,174. Conclusions Mild HSE may not present with classic frontotemporal lobe syndrome and fever may not be an inevitable symptom in patients with immunosuppression. However, the possibility of HSE should be considered in patients with atypical intracranial infection, and these patients should be tested by NGS.

scholarly journals Disseminated Strongyloides Stercoralis Hyperinfection in An Immunocompetent Patient First Diagnosed by Metagenomic Next-Generation Sequencing in Cerebrospinal Fluid: A Case Report

2021 ◽  
Author(s):  
Junyan Qu ◽  
Zhiyong Zong
Keyword(s):  
Cerebrospinal Fluid ◽  
Abdominal Pain ◽  
Next Generation Sequencing ◽  
Clinical Manifestations ◽  
Good Health ◽  
Strongyloides Stercoralis ◽  
Next Generation ◽  
Diagnostic Dilemma ◽  
Disseminated Strongyloidiasis ◽  
Generation Sequencing

Abstract Background Disseminated Strongyloides stercoralis hyperinfection is rarely described in immunocompetent individuals and can lead to fatal outcomes if not recognized and diagnosed early. Non-specific clinical manifestations, such as pneumonia and gastroenteritis, pose a diagnostic dilemma. Case presentation: We report a case of a 67-year-old Chinese male who presented with two months of abdominal pain, fever, headache, vomiting, constipation, and slight cough with sputum. He had been in good health and had no history of glucocorticoid use. He was diagnosed with enterococcal meningitis and intestinal obstruction at a local hospital and improved after treatment with vancomycin, but symptoms of headache and abdominal pain soon recurred. The metagenomic next-generation sequencing (mNGS) of the cerebrospinal fluid using Illumina X10 sequencer revealed 7 sequence reads matching Strongyloides stercoralis. Disseminated strongyloidiasis was suspected. Next, microscopic examination of gastric fluid revealed Larvae of S. stercoralis. DNA extracted of larvae, the presence of both S. stercoralis ribosomal DNA gene and mitochondrial cytochrome c oxidase subunit 1 gene was identified. Disseminated strongyloidiasis was diagnosed. Albendazole (400 mg, twice daily) was used and the patient recovered gradually. Conclusions S. stercoralis hyperinfection can occur in immunocompetent individuals, imposing challenges for diagnosis. mNGS may be a useful tool for detecting rare infectious disease. The case would help clinicians to raise awareness of strongyloidiasis in non-endemic areas and reduce fatality.

scholarly journals Metagenomic Next Generation Sequencing in the Detection of Pathogens in Cerebrospinal Fluid of Patients After Alternative Donor Transplantation: A Feasibility Analysis

2021 ◽  
Vol 11 ◽  
Author(s):  
Binglei Zhang ◽  
Jian Zhou ◽  
Ruirui Gui ◽  
Zhen Li ◽  
Yingling Zu ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Next Generation Sequencing ◽  
Viral Infections ◽  
Fungal Infections ◽  
Clinical Manifestations ◽  
Next Generation ◽  
Hematopoietic Stem ◽  
Cns Infections ◽  
Alternative Donor ◽  
Generation Sequencing

Central nervous system (CNS) complications can occur in 9%–15% of patients after allogeneic hematopoietic stem cell transplantation (allo-HSCT). The clinical manifestations of the CNS complications are non-specific, with most of them being disturbances of consciousness, convulsions, headaches, fever, and epilepsy, making it difficult to infer the cause of the complications based on clinical manifestations. We retrospectively analyzed the sensitivity and feasibility of metagenomic next generation sequencing (mNGS) in the diagnosis of CNS infections after allo-HSCT. Lumbar punctures were performed on 20 patients with CNS symptoms after receiving alternative donor HSCT(AD-HSCT) at the Affiliated Cancer Hospital of Zhengzhou University from February 2019 to December 2020, and their cerebrospinal fluid (CSF) was collected. The mNGS technique was used to detect pathogens in the CSF. Routine CSF testing, biochemical analyses, G experiments, GM experiments, ink staining, acid-fast staining, and bacterial cultures were carried out, and quantitative PCR (qPCR) tests were used to detect cytomegalovirus (CMV), Epstein-Barr virus (EBV), BK polyomavirus (BKPyV), and human alphaherpesvirus (HHV). A total of 29 tests were performed with 21 of them being positive. Of the five negative patients, three were diagnosed with a posterior reversible encephalopathy syndrome, one as having transplantation-associated thrombotic microangiopathy, and one with transient seizure caused by hypertension. Fifteen patients tested positive, of which four had single infections and eleven had mixed infections. Five cases of fungal infections, six cases of bacterial infections, and 13 cases of viral infections were detected. Among the 13 cases of viral infections, ten cases were CMV(HHV-5); three were BKPyV; two were Torque teno virus (TTV); Two were HHV-1,two were EBV(HHV4), and one each of HpyV5 and HHV-6B. Thirteen patients tested positive for virus while the qPCR detection method of 6 identical specimens were below the minimum detection limit(<1×103 U/ml). The mNGS technique is highly sensitive, and it can be used to diagnose CNS infections after allo-HSCT.

scholarly journals 1224Defining herpes simplex virus type 2 sequence heterogeneity using next generation sequencing

2014 ◽  
Vol 1 (suppl_1) ◽  
pp. S42-S43
Author(s):  
Christine Johnston ◽  
Amalia Magaret ◽  
Kurt Diem ◽  
Matthew Fitzgibbon ◽  
Meei-Li Huang ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Next Generation Sequencing ◽  
Herpes Simplex ◽  
Virus Type ◽  
Herpes Simplex Virus Type ◽  
Next Generation ◽  
Sequence Heterogeneity ◽  
Simplex Virus ◽  
Generation Sequencing

Next generation sequencing identifies multi-drug resistant herpes simplex virus- associated scrotal ulceration

2020 ◽  
Vol 80 (2) ◽  
pp. 232-254 ◽  
Author(s):  
Yin Mo ◽  
Chun Kiat Lee ◽  
Tze Ping Loh ◽  
Evelyn Siew Chuan Koay ◽  
Julian W. Tang ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Next Generation Sequencing ◽  
Herpes Simplex ◽  
Drug Resistant ◽  
Next Generation ◽  
Simplex Virus ◽  
Generation Sequencing

scholarly journals Disseminated Strongyloides Stercoralis Hyperinfection in an Immunocompetent Patient First Diagnosed by Metagenomic Next-generation Sequencing in Cerebrospinal Fluid: a Case Report

2021 ◽  
Author(s):  
Junyan Qu ◽  
Zhiyong Zong
Keyword(s):  
Cerebrospinal Fluid ◽  
Abdominal Pain ◽  
Next Generation Sequencing ◽  
Clinical Manifestations ◽  
Good Health ◽  
Strongyloides Stercoralis ◽  
Next Generation ◽  
Diagnostic Dilemma ◽  
Disseminated Strongyloidiasis ◽  
Generation Sequencing

Abstract Background: Disseminated Strongyloides stercoralis hyperinfection is rarely described in immunocompetent individuals and can lead to fatal outcomes if not recognized and diagnosed early. Non-specific clinical manifestations, such as pneumonia and gastroenteritis, pose a diagnostic dilemma. Case presentation: We report a case of a 67-year-old Chinese male who presented with two months of abdominal pain, fever, headache, vomiting, constipation, and slight cough with sputum. He had been in good health and had no history of glucocorticoid use. He was diagnosed with enterococcal meningitis and intestinal obstruction at a local hospital and improved after treatment with vancomycin, but symptoms of headache and abdominal pain soon recurred. The metagenomic next-generation sequencing (mNGS) of the cerebrospinal fluid using Illumina X10 sequencer revealed 7 sequence reads matching Strongyloides stercoralis. Disseminated strongyloidiasis was suspected. Next, microscopic examination of gastric fluid revealed Larvae of S. stercoralis. DNA extracted of larvae, the presence of both S. stercoralis ribosomal DNA gene and mitochondrial cytochrome c oxidase subunit 1 gene was identified. Disseminated strongyloidiasis was diagnosed. Albendazole (400 mg, twice daily) was used and the patient recovered gradually.Conclusions: S. stercoralis hyperinfection can occur in immunocompetent individuals, imposing challenges for diagnosis. mNGS may be a useful tool for detecting rare infectious disease. The case would help clinicians to raise awareness of strongyloidiasis in non-endemic areas and reduce fatality.

scholarly journals The metagenomic next-generation sequencing in diagnosing central nervous system angiostrongyliasis: a case report

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Li Feng ◽  
Aiwu Zhang ◽  
Jiali Que ◽  
Hongyan Zhou ◽  
Haiyan Wang ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Next Generation Sequencing ◽  
Clinical Manifestations ◽  
Next Generation ◽  
Opening Pressure ◽  
Csf Analysis ◽  
Elevated Protein ◽  
The Central Nervous System ◽  
Generation Sequencing

Abstract Backgrounds The incidence of angiostrongyliasis is increasing in recent decades due to the expanding endemic areas all over the world. Clinicians face tremendous challenge of diagnosing angiostrongyliasis because of the lack of awareness of the disease and less effective definitive laboratory tests. Case presentation A 27-year-old man initially manifested skin itching, emesis, myalgia and quadriparesis. With progressive weakness of four limbs and elevated protein in the cerebrospinal fluid (CSF), he was diagnosed as Guillain-Barré syndrome and treated with intravenous methylprednisolone and immunoglobulin. However, the patient deteriorated with hyperpyrexia, headache and then persistent coma. The routine tests for Angiostrongylus cantonensis (A. cantonensis) with both the CSF and the serum were all negative. In contrast, the metagenomic next-generation sequencing (mNGS) was applied with the serum sample and the CSF sample in the middle phase. The central nervous system (CNS) angiostrongyliasis was diagnosed by mNGS with the mid-phase CSF, but not the mid-phase serum. At the same time, the CSF analysis revealed eosinophils ratio up to 67%. The discovery of A. cantonensis was confirmed by PCR with CSF later. Unfortunately, the patient died of severe angiostrongyliasis. During his hospitalization, mNGS was carried out repeatedly after definitive diagnosis and targeted treatment. The DNA strictly map reads number of A. cantonensis detected by mNGS was positively correlated with the CSF opening pressure and clinical manifestations. Conclusions The case of A. cantonensis infection highlights the benefit of mNGS as a target-free identification in disclosing the rare CNS angiostrongyliasis in the unusual season, while solid evidence from routine clinical testing was absent. The appropriate sample of mNGS should be chosen according to the life cycle of A. cantonensis. Besides, given the fact that the DNA reads number of A. cantonensis fluctuated with CSF opening pressure and clinical manifestations, whether mNGS could be applied as a marker of effectiveness of treatment is worth further exploration.

scholarly journals Development and Optimization of Metagenomic Next-Generation Sequencing Methods for Cerebrospinal Fluid Diagnostics

2018 ◽  
Vol 56 (9) ◽  
Author(s):  
Patricia J. Simner ◽  
Heather B. Miller ◽  
Florian P. Breitwieser ◽  
Gabriel Pinilla Monsalve ◽  
Carlos A. Pardo ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Next Generation Sequencing ◽  
Sample Pretreatment ◽  
Standard Of Care ◽  
Nucleic Acid Amplification ◽  
Next Generation ◽  
Bead Beating ◽  
Dna And Rna ◽  
Positive Threshold ◽  
Generation Sequencing

ABSTRACT The purpose of this study was to develop and optimize different processing, extraction, amplification, and sequencing methods for metagenomic next-generation sequencing (mNGS) of cerebrospinal fluid (CSF) specimens. We applied mNGS to 10 CSF samples with known standard-of-care testing (SoC) results (8 positive and 2 negative). Each sample was subjected to nine different methods by varying the sample processing protocols (supernatant, pellet, neat CSF), sample pretreatment (with or without bead beating), and the requirement of nucleic acid amplification steps using DNA sequencing (DNASeq) (with or without whole-genome amplification [WGA]) and RNA sequencing (RNASeq) methods. Negative extraction controls (NECs) were used for each method variation (4/CSF sample). Host depletion (HD) was performed on a subset of samples. We correctly determined the pathogen in 7 of 8 positive samples by mNGS compared to SoC. The two negative samples were correctly interpreted as negative. The processing protocol applied to neat CSF specimens was found to be the most successful technique for all pathogen types. While bead beating introduced bias, we found it increased the detection yield of certain organism groups. WGA prior to DNASeq was beneficial for defining pathogens at the positive threshold, and a combined DNA and RNA approach yielded results with a higher confidence when detected by both methods. HD was required for detection of a low-level-positive enterovirus sample. We demonstrate that NECs are required for interpretation of these complex results and that it is important to understand the common contaminants introduced during mNGS. Optimizing mNGS requires the use of a combination of techniques to achieve the most sensitive, agnostic approach that nonetheless may be less sensitive than SoC tools.

scholarly journals Diagnosis of Enterococcus faecalis meningitis associated with long-term cerebrospinal fluid rhinorrhoea using metagenomics next-generation sequencing: a case report

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Xiaobo Zhang ◽  
Chao Jiang ◽  
Chaojun Zhou
Keyword(s):  
Cerebrospinal Fluid ◽  
Next Generation Sequencing ◽  
Enterococcus Faecalis ◽  
Bone Defect ◽  
Next Generation ◽  
History Of ◽  
Diagnostic Approaches ◽  
Post Traumatic ◽  
Skull Bone Defect ◽  
Generation Sequencing

Abstract Background Enterococcus faecalis (E. faecalis) meningitis is a rare disease, and most of its occurrences are of post-operative origin. Its rapid diagnosis is critical for effective clinical management. Currently, the diagnosis is focused on cerebrospinal fluid (CSF) culture, but this is quite limited. By comparison, metagenomic next-generation sequencing (mNGS) can overcome the deficiencies of conventional diagnostic approaches. To our knowledge, mNGS analysis of the CSF in the diagnosis of E. faecalis meningitis has been not reported. Case presentation We report the case of E. faecalis meningitis in a 70-year-old female patient without a preceding history of head injury or surgery, but with an occult sphenoid sinus bone defect. Enterococcus faecalis meningitis was diagnosed using mNGS of CSF, and she recovered satisfactorily following treatment with appropriate antibiotics and surgical repair of the skull bone defect. Conclusions Non-post-traumatic or post-surgical E. faecalis meningitis can occur in the presence of occult defects in the cranium, and mNGS technology could be helpful in diagnosis in the absence of a positive CSF culture.

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