Validation of Metagenomic Next-Generation Sequencing Tests for Universal Pathogen Detection.

Context.--Metagenomic sequencing can be used for detection of any pathogens using unbiased, shotgun nextgeneration sequencing (NGS), without the need for sequence-specific amplification. Proof-of-concept has been demonstrated in infectious disease outbreaks of unknown causes and in patients with sus...

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Publicado en:Archives of Pathology & Laboratory Medicine Vol. 141; no. 6; pp. 776 - 787
Autores principales: Schlaberg, Robert, Chiu, Charles Y., Miller, Steve, Procop, Gary W., Weinstock, George
Formato: research tables/charts Journal Article
Publicado: College of American Pathologists Jun2017
Acceso en línea:Ver este registro en EBSCOhost
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      dt: Jun2017
      vid: 141
      iid: 6
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      pub: College of American Pathologists
      place: Northfield, Illinois
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        10.5858/arpa.2016-0539-RA
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        atl: Validation of Metagenomic Next-Generation Sequencing Tests for Universal Pathogen Detection.
      aug:
        au:
          Schlaberg, Robert
          Chiu, Charles Y.
          Miller, Steve
          Procop, Gary W.
          Weinstock, George
        affil: Department of Pathology, University of Utah, and the Institute for Clinical and Experimental Pathology, ARUP Laboratories, Salt Lake City, Utah
      sug:
        subj:
          Genome
          Infection Diagnosis
          Sequence Analysis
          Pathology
          Validation Studies
          Communicable Diseases
          Genetics
          Oncology
          Data Collection
          Data Analysis Software
          Biological Assay
          Virus Diseases Diagnosis
          Bacterial Infections
          Mycoses
          Polymerase Chain Reaction
          Medical Organizations
          Cerebrospinal Fluid Physiology
          Sensitivity and Specificity
          Reproducibility of Results
          Databases
          Specimen Handling
      ab: Context.--Metagenomic sequencing can be used for detection of any pathogens using unbiased, shotgun nextgeneration sequencing (NGS), without the need for sequence-specific amplification. Proof-of-concept has been demonstrated in infectious disease outbreaks of unknown causes and in patients with suspected infections but negative results for conventional tests. Metagenomic NGS tests hold great promise to improve infectious disease diagnostics, especially in immunocompromised and critically ill patients. Objective.--To discuss challenges and provide example solutions for validating metagenomic pathogen detection tests in clinical laboratories. A summary of current regulatory requirements, largely based on prior guidance for NGS testing in constitutional genetics and oncology, is provided. Data Sources.--Examples from 2 separate validation studies are provided for steps from assay design, and validation of wet bench and bioinformatics protocols, to quality control and assurance. Conclusions.--Although laboratory and data analysis workflows are still complex, metagenomic NGS tests for infectious diseases are increasingly being validated in clinical laboratories. Many parallels exist to NGS tests in other fields. Nevertheless, specimen preparation, rapidly evolving data analysis algorithms, and incomplete reference sequence databases are idiosyncratic to the field of microbiology and often overlooked.
      pubtype: Academic Journal
      doctype:
        research
        tables/charts
        Journal Article
      ougenre: Article
    language: English
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