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Report from the Standards for Pathogen Identification via Next-Generation Sequencing (SPIN) Workshop


Next-generation sequencing (NGS) is not routinely used in applied settings due to lack of confidence in results. This workshop convened experts to identify measurement challenges impeding NGS implementation and potential standards-based solutions to address these challenges.

Sequencing of microbial genomes, a task that once took large sequencing centers months to complete, is regularly performed by small laboratories in a few days due to the development of benchtop next-generation sequencing (NGS) platforms. This capability has resulted in a desire to use whole genome sequencing (WGS) for pathogen identification in applied settings such as biothreat detection, molecular epidemiology, and clinical diagnostics. However, a number of challenges related to sample processing and data analysis must be overcome before widespread adoption of WGS is realized.

In an effort to identify priority areas for standards activities and facilitate the development of a measurement infrastructure for NGS-based pathogen identification, the National Institute of Standards and Technology (NIST) convened a two-day workshop composed of representatives from Federal Government, academia, and industry. The workshop took place on October 20–21, 2014 at the NIST campus in Gaithersburg, Maryland. The objectives of the Standards for Pathogen Identification via Next-Generation Sequencing (SPIN) Workshop were to identify current and anticipated future measurement challenges hindering the implementation of NGS in pathogen identification, and to propose avenues to address these challenges including recommendations for standards development (see NIST Special Publication SP1183 for a detailed description of the workshop including summaries of the presentations) [1].

On the first day of the workshop, leaders in the field presented their efforts and thoughts on WGS challenges related to specific areas including metrology, sample preparation, molecular epidemiology, antimicrobial resistance surveillance, large-scale genome sequencing projects, genome sequence databases, bioinformatics methods, and biomarker development (Table 1). These application areas included strain-level isolate identification and discrimination as well as culture-independent diagnostics from complex samples. (The slides for a number of the presentations are available online [2]). Despite the diversity of topics, several recurring themes were identified including:

  • the need for standard methods and reference materials for sample processing and data analysis,

  • performance metrics for validating data analysis methods,

  • well curated and diverse databases, and

  • guidance on results interpretation.

Table 1 Workshop speakers and presentation titles

The majority of the second day consisted of small and large group discussions to identify current and future measurement challenges associated with implementation of NGS for pathogen identification and potential solutions for these challenges. The entire process, from sample to answer, is challenging and requires a supporting measurement infrastructure to increase confidence in the final results. Each step in the process has measurement challenges, biases, uncertainties, and application-specific components, requiring measurement solutions and standards. Table 2 describes four primary classes of solutions often used to address measurement challenges.

Table 2 Classes of metrology-based solutions to support a measurement infrastructure

During the workshop, several specific examples of standards-based solutions were discussed. Some of these solutions were identified as potential next steps for standards development based upon their anticipated high impact, expected usage, and relative ease of preparation as compared to other activities. This list is not exhaustive but serves to identify examples related to the four categories of measurement-based solutions.

  • Reference data: Workshop attendees discussed that development of in silico reference data might be a simpler starting point than physical reference materials (such as cells or DNA). Datasets could be created by combining known genome sequences and used to benchmark and compare bioinformatics pipelines and results reporting and interpretation. These datasets could include relevant pathogens as well as environmental contaminants, such as host DNA, other microorganisms, and viral genomes.

  • Reference materials: Well-characterized microbial genomic DNA reference materials are needed for validation of sequencing platforms, library preparation protocols, sequencing chemistries, and base calling algorithms.

  • Guidance documents: Reference materials specific for each application area are needed, yet development of such a vast set of reference materials is too large for any one organization. Guidance documents describing methods to develop and characterize control materials would enable industry and others to prepare quality control materials (in house development of the materials) with organisms relevant to their specific application space.

  • Interlaboratory studies: An interlaboratory study to evaluate the contents of a known mixture of well-characterized microbes is needed to support metagenomic analyses. Results from the study would be used to characterize sources of bias and uncertainty associated with metagenomic sequencing, including DNA extraction, sample processing, and data analysis.

The first steps in establishing a measurement infrastructure for pathogen identification using NGS are already underway. For instance, proposals for documentary standards related to NGS are under consideration by International Organization for Standardization (ISO) TC34: Food Products. These proposals include a standard for foodborne pathogen strain typing using WGS and a more general standard on NGS quality analysis. In a separate effort, NIST in collaboration with the FDA is developing genomic DNA reference materials for four bacterial strains, Salmonella enterica subsp. enterica serovar Typhimurium strain LT2, Staphylococcus aureus clinical isolate, Pseudomonas aeruginosa clinical isolate, and Clostridium sporogenes PA3679. These materials will be used to validate sequencing chemistries and platforms and to support laboratory proficiency testing. Additionally, data generated from the material can be used to validate bioinformatics workflows such as de-novo assembly and variant calling.

Overall, the solutions to the challenges identified during the workshop will serve as the basis for a measurement infrastructure for pathogen identification using WGS. NIST is in a unique position to help advance this field by providing expertise in metrology (measurement science) and by leveraging experience developing reference materials and measurement infrastructures for related fields, such as human genome sequencing [3] and transcriptome sequencing [4]. A similar measurement infrastructure for pathogen identification using NGS will increase confidence in results and improve data informed decision-making, in turn enabling WGS to achieve its full potential in revolutionizing pathogen identification.



Next Generation Sequencing


Whole Genome Sequencing


National Institute of Standards and Technology


International Organization for Standards


Deoxyribonucleic Acids


Food and Drug Administration


National Center for Biotechnology Information


  1. 1.

    Olson ND, Jackson SA, and Lin NJ. Standards for Pathogen Identification via Next-Generation Sequencing (SPIN) Workshop Summary Report. NIST Special Publication 1183

  2. 2.

    Publically available presentations from the workshop. Accessed 25 October 2015.

  3. 3.

    Genome In A Bottle. Accessed 25 October 2015.

  4. 4.

    External RNA Control Consortium. Accessed 25 October 2015.

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We would like to thank all the workshop presenters as well as those who helped run the workshop: Sandra Da Silva, Katherine Gettings, Samuel Forry, Arlin Stoltzfus, William Wolfgang, Robert (Chris) Hopkins, and Bart Weimer. The SPIN Workshop was hosted by NIST and funded by the Department of Homeland Security (DHS) Science and Technology Directorate under Interagency Agreement HSHQPM-13-X-00190 with NIST. Opinions expressed in this report are the authors’ and do not necessarily reflect the policies and views of DHS, NIST, or affiliated venues. Official contribution of NIST; not xsubject to copyrights in USA.

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Corresponding author

Correspondence to Nathan D. Olson.

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Competing interests

The authors declare that they have no competing interests.

Authors’ contributions

NDO, SAJ, and NJL all contributed to drafting the meeting report. All authors read and approved the final manuscript.

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Olson, N.D., Jackson, S.A. & Lin, N.J. Report from the Standards for Pathogen Identification via Next-Generation Sequencing (SPIN) Workshop. Stand in Genomic Sci 10, 119 (2015).

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  • Genome sequencing
  • Metrology
  • Next-generation sequencing
  • Pathogen identification
  • Reference materials
  • Standards