microbial diagnostics - university of calgary in alberta
TRANSCRIPT
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Ian Lewis, PhD; Assistant ProfessorAI Translational Health Chair
University of Calgary
www.lewisresearchgroup.org
Microbial diagnostics
AMR Course | Calgary, AB | 2020
Lewis Research Group
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Antibiotic resistance in Calgary
Drug sensitive
Drug Resistant
Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
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June 19th
18:15
June 24st
10:01
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Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
Youtube video on microbiology testing
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Emerging platforms for fast ID/AST
Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
Direct MALDI-TOF mass spectrometry
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Emerging platforms for fast ID/AST
Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
Biofire film array (BCID panel) (Multiplex real-time PCR)
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Emerging platforms for fast ID/AST
Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
216DX UTI System (Laser light scattering)
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Emerging platforms for fast ID/AST
Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
T2 Candida Panel (T2 Magnetic Resonance)
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Emerging platforms for fast ID/AST
Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
Reveal AST (Volatile organic compound detection)
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Emerging platforms for fast ID/AST
Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
AStar (microscopy)
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Emerging platforms for fast ID/AST
Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
Phoenix Automated Microbiology System (Redox and Turbidity)
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Kumar A et al. Crit Care Med 2006; 34:1589-96.
Deaths resulting from slow diagnostics
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Can metabolism be used as a diagnostic platform?
Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
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Can metabolism be used as a diagnostic platform?
Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
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Calgary Metabolomics Research Facility (CMRF)
Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
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Thomas Rydzak, PhDUniversity of Calgary
Metabolic Preference Assay
Apex/DynaLIFE NDA
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Can microbes be identified via metabolism?
YesN = 23,000
Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
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Can antibiotic susceptibility can be measured by metabolism?
Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
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Yes, metabolite profiles are highly diagnostic
Can antibiotic susceptibility can be measured by metabolism?
Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
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Does it work? A blinded trial of Alberta
1. Every blood culture in Calgary
2. 10 days of collection
3. N = 775
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Sensitivity (%) Specificity (%)
Culture growth 96.1 99.8
Genus ID 84 87
n=775Species ID of 7 target species = 85.8%
Did it work? Yes!!!
negative
positive
Samples containing organisms not within training set Samples containing organisms within training set
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The BIG Race
Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
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Tom wins!!!!!!
Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
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Why UTIs are a problem
• Most common form of bacterial infection
• Estimated cost of $2.6B USD annually
• Potentially life-threatening consequences
• Lifetime occurrence of 50-70% in women
Dan Gregson, MDCalgary Laboratory Services
Long diagnostic timelines: a familiar problem
Can metabolomics detect UTIs?
Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
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Can metabolomics detect UTIs?
Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
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Can metabolomics detect UTIs?
Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
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METHOD DEVELOPMENTMETHOD
1. Urine collected 2. 13C labeled standard added.
3. Target metabolites are isolated
4. LC-MS SRM analysis (<1min per sample)
Rapid screening assay for urinary tract infection
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Can metabolomics detect UTIs?
Spencer WildmanUniversity of Calgary
Ex1: BSI Test Ex2: UTI TestWhy omics? PIM Innovation
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Tom versus Alberta Public Laboratories
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Microbiome
Lukas F. Mager et al. Science, 2020
Yonatan GradHarvard School of Public Health
George ChaconasUniversity of Calgary
Kathy McCoyUniversity of Calgary
Steven NorrisUniversity of
Texas
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Rapid Infection Diagnostics Inc.
HOPPER – BSIDXTM and UTIDXTM test
Transwell with
engineered medium
and ABs
Vanquish sample hotel and
UHPLC
TSQ Altis Triple Quad ID database
What is APEX?Why APEX? APEX’s PotentialExample Project Achievements InnovationAchievementsEx1: BSI Test Ex2: UTI TestWhy Omics? PIM
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Rapid diagnostics for automatable surveillance
What is APEX?Why APEX? APEX’s PotentialExample Project Achievements InnovationAchievementsEx1: BSI Test Ex2: UTI TestWhy Omics? PIM
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AMR Surveillance
Simon Otto, PhD DVM BSc
Assistant Professor
We acknowledge and pay tribute to the traditional territories of the peoples of Treaty 6, which includes the Cree, Blackfoot, Métis, Nakota Sioux, Iroquois, Dene, Ojibway, Saulteaux, Anishinaabe, Inuit, and many others
whose histories, languages, and cultures continue to influence our vibrant community.
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Outline
1. Intro: Dr. Ian Lewis and Dr. Simon Otto
2. Integrated AMR/AMU surveillance
3. Dr. Lewis: AMR measurement and related
sections
4. Whole Genome Sequencing (WGS) for
AMR surveillance
5. Select AMR/AMU Surveillance Examples
6. Discussion
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Links to Course Learning
Outcomes
• Course Learning Outcomes:1. Complex contributing factors to AMU and the
emergence and spread of AMR.
3. The global interdependence of people, animals,
and the environment (One Health)
4. The role of One Health in complex problems
and AMR.
5. AMR surveillance requirements and mitigation
efforts.
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Learning Objectives (Simon)
• Objective 1: Explain the components of an integrated,
One Health surveillance system for AMR and AMU.
• Objective 2: Describe the opportunities and challenges
of using Whole Genome Sequencing for AMR
surveillance.
• Objective 3: Explain key examples of the application of
integrated, One Health AMR/AMU surveillance.
• Objective 4: Assess the gaps in One Health AMR/AMU
surveillance.
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Be thinking about…
• What does integration in an AMR/AMU
surveillance system entail?
– What must be integrated?
– Why is integration important to inform
stakeholders?
• Practitioners
• Industry
• Government and other policy makers
• Etc.
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Resources• Required resource:
– Miazga-Rodriguez M, Saxinger LM, Otto SJG. “How are we
doing? Progress on Integrated AMR and AMU Surveillance in
Canada: 2014-2019.”
• CPHA Public Health 2020 virtual abstract: https://youtu.be/PD7wGeA8hKU
• Supplemental reading:
– WHO AGISAR 2017 “Integrated surveillance of AMR in
foodborne bacteria Application of a One Health approach”
• https://www.who.int/foodsafety/publications/agisar_guidance2017/en/
– WHO 2020 “Global Antimicrobial Surveillance System - Whole-
genome sequencing for surveillance of AMR”
• https://www.who.int/publications/i/item/9789240011007
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8http://www.phac-aspc.gc.ca/cipars-picra/gfx/epi-lg-eng.png
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What is Surveillance?
• What is surveillance?
– “Information for Action” – Larry Svenson, AH
• What is monitoring?
– Surveillance without an action/response component
• “Systematic ongoing collection, collation, and analysis of
data and the timely dissemination of information to those
who need to know so that action can be taken.” - WHO
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Purpose of AMR Surveillance
• AMR surveillance – “characterization AMR
pathogens and their distribution in the
population” – WHO 2020 GLASS WGS for AMR surveillance
• What is missing from this?
– What population(s)?
– Pathogens are not the only important bugs
• Commensals, indicator organisms, microbiome
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What are the objectives of AMR
surveillance?• Analysis of trends in AMR rates
• Frequency of AMR infections
• Impact of AMR infections on human
health?
• Data to inform antimicrobial categorization
and/or prudent use guidelines/systems
• Data for risk assessment/analysis
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What are the objectives of AMU
surveillance?• Analysis of trends in AMU rates
• Data for risk assessment/analysis
• Ideally linked to AMR data in some
manner
– Level of the population? Individual?
– Goal: understand selection pressure for AMR
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AMU Surveillance Objectives
• Where does this get complicated?
– Linking animal AMU to human AMR
– Linking individual AMU to individual/
population AMR
– Complicated drug distribution system
• Humans but especially animals
– What will you do with the data?
• Educate? Policy? Regulate use? (eg yellow card)
• At what level of the population? Census vs sample
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Integrated AMR/AMU Surveillance?
• What does “integrated” mean?
• For AMR and AMU surveillance,
integration means that data from multiple
sources/populations are analyzed to
understand how AMU affects AMR in the
complex “confusogram” system under
consideration
Many different possible “levels”
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Integrated AMR/AMU Surveillance?
• Sample sources (populations):
– Human specimens
– Food-animals
– Retail food
– Environment? Crops? Water? Soil?
• What is the objective of the program?
– Foodborne transmission?
– Nosocomial infection?
– Information? Education? Regulation?
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Target bacteria?
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Taken from the PHAC CARSS 2018 Stakeholder presentation – CARSS report 2017.
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AMU Surveillance?
• Where, how, whither…?
• AMU data collection is complicated
• Level of collection depends on the level of
comparison
• Integration can take many forms
• There are many ways to measure AMU
– Total kg, mg/PCU, DDDvetCA/1000 ADR…
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AMR/AMU Surveillance
• Integrated AMR/AMU surveillance– This gets very granular very quickly…
• 2014 NCCID report – recommendations for Canada for
integrated surveillance– https://nccid.ca/publications/surveillance-of-antimicrobial-resistance-and-
antimicrobial-utilization-in-canada/
• 2016 CCVO report – AMU surveillance in animals– https://www.cahss.ca/media/uploads/cipars/documents/17-08-01_19-
44/CCVO_AMUCommittee_Non-HumanAMUSurveillance_FINAL_NjgJxTA.pdf
• Update on Canadian progress – CPHA Public Health 2020,
abstract (Miazga-Rodriguez/Saxinger/Otto)– https://youtu.be/PD7wGeA8hKU
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AMR and AMU Surveillance
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https://www.canada.ca
/content/dam/hc-
sc/documents/service
s/drugs-health-
products/canadian-
antimicrobial-
resistance-
surveillance-system-
2020-report/CARSS-
2020-report-2020-
eng.pdf
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Dr. Ian Lewis
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Whole Genome SequencingWHO 2020 WGS: https://www.who.int/publications/i/item/9789240011007
• AMR surveillance still largely depends on
phenotypic testing
• Opportunities for metabolomics (Dr. Lewis)
• Opportunities for genomics and
metagenomics
• In silico and machine learning methods to
infer phenotype from genotype
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WGS Opportunities
• WGS – DNA sequence compared to
reference database
– Identify AMR genes
– Possibly identify virulence genes, promoters,
etc
– Reconstruction of transmission networks
based on molecular (genetic) subtypes
• Good from a One Health perspective
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WGS Opportunities
• Identify molecular determinants and
mechanisms of AMR
• Identify genetic factors facilitating
transmission of AMR genes within microbial
populations
• Useful addition to phenotypic at present
• Metagenomics – avoid the need for pure
culture = less time
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WGS Limitations
• WGS still requires a pure culture of an
organism
• WGS cannot completely substitute
phenotypic methods
– Genotype is not always = phenotype
Role for metabolomics
• Lack of international standards
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Metagenomics Limitations
• Long-read sequencing methods (next-
generation sequencing)
• Methods still not there, but improving!
– Error rates
– Bioinformatics
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Select Surveillance Examples
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Taken from the PHAC CARSS 2016 Stakeholder presentation – CARSS report 2016.
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30Taken from the CIPARS 2019 Stakeholder presentation – integrated 2018 results.
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31Taken from the CIPARS 2019 Stakeholder presentation – integrated 2018 results.
PCU (Population correction units) = biomass of animals in the population↑ 6%
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National comparisons of AMU:
Canada vs EU (mg/PCU)
Assumes that data are comparable
Taken from the CIPARS 2019 Stakeholder presentation – integrated 2018 results.
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Taken from the CIPARS 2017 Stakeholder presentation – integrated 2016 results. 33
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Taken from the CIPARS 2017 Stakeholder presentation – integrated 2016 results.
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Ceftriaxone-resistance in NTS
35Taken from the CIPARS 2019 Stakeholder presentation – integrated 2018 results.
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Highly Drug-resistant Salmonella
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Taken from the CIPARS 2017 Stakeholder presentation – integrated 2016 results.37
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Taken from the CIPARS 2017 Stakeholder presentation – integrated 2016 results.
Co-selection gentamicin + lincomycin-spectinomycin useVeterinary Microbiology 203 (2017) 149–157
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http://www.bccdc.ca/health-professionals/data-reports/antimicrobial-resistance-utilization
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http://www.bccdc.ca/health-professionals/data-reports/antimicrobial-resistance-utilization/antimicrobial-resistance-dashboard
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http://www.bccdc.ca/health-professionals/data-reports/antimicrobial-resistance-utilization/antimicrobial-utilization-dashboard
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Summary
• AMR and AMU surveillance must be
integrated to truly understand the links
between use and resistance
– This integration can be at different levels and
in different ways
– Surveillance program design depends on its
objectives
• There are One Health gaps in surveillance
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Thank you!
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