National Center for Immunization & Respiratory Diseases
Rapid Diagnostic Tests for Bacterial Meningitis Pathogens: where we are now and what’s next.
Xin Wang Chief, Bacterial Meningitis Laboratory Director WHO Collaborating Center for Meningitis MVPDB/DBD/NCIRD/CDC Meningitis Research Foundation Conference Nov 1-3, 2021
The document discusses the current state of rapid diagnostic tests for bacterial meningitis pathogens and outlines a vision for their future development and deployment. It describes existing tests and their limitations. Potential new platforms are identified that could meet targets outlined in a target product profile. Advanced technologies like sequencing and CRISPR/Cas are also discussed
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https://www.meningitis.org/mrf-conference-2017
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https://www.meningitis.org/mrf-conference-2017
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Prospects for GBS prevention - current candidates & removing barriers to licensure of a GBS vaccine for pregnant women globally
https://www.meningitis.org/mrf-conference-2017
Single-dose oral ciprofloxacin prophylaxis as a meningococcal meningitis outbreak response: results of a cluster-randomized trial
https://www.meningitis.org/mrf-conference-2017
What next for prevention of pneumococcal disease in light of serotype replacement? Is there a pathway to licensure for novel pneumococcal vaccines?
https://www.meningitis.org/mrf-conference-2017
Best Practices for Validating a Next-Gen Sequencing WorkflowGolden Helix
Validating an NGS workflow is an iterative process that begins with collaboration with personnel and planning protocols for the entire workflow from sample preparation, sequencing and variant calling, all the way to data analysis and reporting. At Golden Helix, while we do not provide pre-validated black-box workflows, we provide our customers with support to validate workflows in a transparent manner, and assist them in reaching production deadlines. This webcast will be led by members of our Field Application Scientist team, and we will explore some of the best practices for NGS workflow validation that we have observed and helped to implement based on real-world examples from our customer base. Key topics for discussion will include:
Sample preparation and collection of adequate case/control data
Designing a robust workflow with special considerations for single versus family analyses and phenotypic considerations
Generating the desired output for clinical or other reports
Real world NGS workflow validation strategies
Tune in for tips and strategies that you can deploy when designing and validating your NGS workflow.
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Earlier this year, we released VarSeq 2.3.0 which brought massive updates to our VSClinical AMP interface, such as enhanced capabilities for automation and analysis of structural variants in the cancer context. Naturally, we wanted to follow that up shortly with similar advancements to our VSClinical ACMG interface, and also make our customers doing germline variant analysis happy.
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VISION
Being proactive
Supporting optimal animal and human health
Exploring ways to reduce overall use of antimicrobials
Using the drugs that prevent and treat disease by killing microscopic organisms in a responsible way
GOAL
to prevent the generation and spread of antimicrobial resistance (AMR). Doing so will preserve the effectiveness of these drugs in animals and humans for years to come.
being to preserve human and animal health and the effectiveness of antimicrobial medications.
to implement a multidisciplinary approach in assembling a stewardship team to include an infectious disease physician, a clinical pharmacist with infectious diseases training, infection preventionist, and a close collaboration with the staff in the clinical microbiology laboratory
to prevent antimicrobial overuse, misuse and abuse.
to minimize the developme
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1. National Center for Immunization & Respiratory Diseases
Rapid Diagnostic Tests for Bacterial
Meningitis Pathogens: where we are now
and what’s next.
Xin Wang
Chief, Bacterial Meningitis Laboratory
Director WHO Collaborating Center for Meningitis
MVPDB/DBD/NCIRD/CDC
Meningitis Research Foundation Conference
Nov 1-3, 2021
2. 2
I have no conflict of interest in relation to this presentation
The findings and conclusions in this presentation are those of the authors and do
not necessarily represent the official position of the U.S. Centers for Disease
Control and Prevention (CDC).
The U.S. CDC does not endorse, recommend or make representations with
respect to the products and manufactures in this presentation
3. Rapid Diagnostics for Meningitis
• Meningitis is a life-threatening disease
• Rapid detection of meningitis pathogens is critical for case management, outbreak
response, and surveillance
• Poor accessibility of diagnostics remains to be addressed, especially in resource
limited countries
• insufficient funding
• limitations of existing diagnostic tests
• lack of trained staff members
• low priority
• Ineffective supply chain management
• Despite advances in diagnostic technology, an empirical antimicrobial treatment
provided, rather than a treatment based on pathogen identification
4. Diagnostics: a Key Component in the Global Road Map to
Defeat Meningitis by 2030
Pillar 2 Diagnosis and treatment
Achieved through improve diagnosis at all levels of heath
care, health worker training and prompt and effective case
management
• Strategic goal 6: Improve diagnosis of meningitis at all levels
of care
• Strategic goal 7: Develop and facilitate access to diagnostic
assays at all levels of care to increase confirmation of
meningitis
Defeating meningitis by 2030: a global road map (who.int)
5. Three Use Cases To Improve Global Meningitis Detection
Use case 1 (Epidemic/outbreak settings, Africa)
• Identification of Nm serogroup at peripheral level (health center or
district hospital) for appropriate vaccine response
Use case 2 (Epidemic and endemic settings, worldwide)
• Identification of bacterial meningitis/septicemia at peripheral level
(health clinic or hospital) to initiate antibiotic treatment for case
management
Use case 3 (Epidemic and endemic settings, worldwide)
• Identification of causative pathogens from syndromic meningitis panels
(minimum 10 pathogens) at hospital level (district/regional hospital) for
case management: stopping or changing antibiotics
https://www.who.int/emergencies/diseases/meningitis/meningitis-diagnostics-use-cases.pdf
6. Timelines for use case 3 development
May 2017: A call
for action: global
vision to defeat
meningitis
March 2018:
Expert group
meeting to
discuss
development of
next generation
RDTs (use cases
1-3)
2019-20: Use
case 3 TPP
finalized
2020-21: Use case
3 landscape and
Market analysis by
PATH
2021: Identification
of potential
platforms for use
case 3
2022-24:
Development and
validation of
potential RDTs
2025-28: Pilot
studies and RDT
roll-out
7. Partnership and collaborations to accelerate
use case 3 development
Develop and finalize
use case 3 Target
Product Profile (TPP)
Conduct landscape
analysis of meningitis
diagnostics
8. Use case 3
TPP
Scope
• Identify the causative meningitis pathogens
• Used in hospital or hospital laboratories
• Performed by trained clinical staff and lab technicians
• Inform appropriate treatment intervention
Specific features
• Multiplex technology allows detection of a wide spectrum of
pathogens
• High performance (sensitivity, specificity, reproducibility, etc)
• Rapid and easy result interpretation
• Ideally, a portable and battery-operated device
• Easy to deploy and use
• Affordable
Meningitis pathogen panel
• Categories A, B and C
• Bacterial, Viral, Fungal, and Parasitic
9. Landscape
analysis of
meningitis
diagnostics
Objectives
• Identify diagnostic gaps and obtain key stakeholders’
feedback on existing and pipeline technologies
• Review the diagnostic platforms and technologies currently
available or under development with the potential for Use
Case 3.
• Assess existing and emerging technologies, including their
quality, cost, and relevance to Use Case 3
Methods
• Stakeholder interviews
• Literature review
Major Findings
• Existing Tests/platforms
• Potential platforms
• Advanced/Emerging technologies
10. Existing Tests and Limitations
Culture
Latex Agglutination
Tests
• Target various meningitis
pathogens and some
meningococcal serogroups
•Rapid (< 20 mins)
•High cost
•Cold chain for
storage/distribution
•Performance may vary
•lab verification: 33-100%
sensitivity; 93-100%
specificity
•field evaluation: 69-80%
sensitivity; 81-94%
specificity
Immunochromatogra
phic tests
• Sp and all meningococcal
serogroups except B
•Rapid (<15 mins)
•Cassette format expensive
•Cold chain for
storage/distribution
•High sensitivity and
specificity for specific
targets
11. 11
• High cost (expensive equipment)
• Require cold chain for key reagents
• Require technical trainings and lab infrastructure
(freezer, fridge, separate rooms etc)
• Decentralization to regional/district levels is
challenging in resource limited regions
• Quick turnaround (within several hours)
• Sensitive/specific for targets
• High throughput
• Multiple platforms available-commercial and
lab developed tests (LDTs)
• LDTs implemented in many countries with
External Quality Assurance in place
Existing Tests and Limitations: PCR-based Tests
12. PCR tests for meningitis pathogens
Test Platform Targets Specimen
Type
Sen/Spe Instrument/Cost Time
BioFire Film
Array
Multiplex Bacterial/viral/fugal meningitis
pathogens
CSF >90% Biofire FilmArray
Systems/High ($45K for
2.0)
~1 hr
Xpert® EV Multiplex Enteroviral meningitis pathogens CSF >95% Genexpert
system/Medium-High
($11K-64K)
<2.5 hrs
QIAstat-Dx Multiplex Bacterial/viral/fugal meningitis
pathogens including Nm/Hi capsule
types
CSF NA QIAStart-Dx
analyzer/Medium
(~$25K)
~1 hr
HG
Meningococcus/
Sp (Ireland)
LAMP Nm Serogroups (A, B, C, E, W, X, Y, and
Z)/Sp serotypes
Blood, CSF,
Swab, Direct
CSF
NA LAMP instrument/$9.7
(main or battery power)
<1 hr
Lab Developed
Tests
Direct PCR,
triplex
Bacterial meningitis pathogens and
capsule types
CSF, serum >95% ABI, AriaMx (>25K) ~ 2 hrs
13. Potential diagnostic platforms
(for other pathogens)
13
Platform Feature Run Time Power Instrument cost
Q-POC Multiplex, up to
40 targets
< 30 mins Main/battery
power
$28K
Anitoa Maverick
compack qPCR
Multiplex, up to 4
targets
~30 mins Main/battery
power
~3.5K-6K
VERI-Q PCR Multiplex, up to
10 targets
< 1 hr Main power ~10K
14. Advanced diagnostic
platforms
• Growing interest in next generation sequencing in past
10 years
• Used for detection of various pathogens (bacterial, viral
etc)
• Various platforms (MinION, Illumin supported platforms,
Ion Torrent, BGISEQ etc) and analysis tools available
• Higher cost and lower sensitive compared to PCR-based
tests
• Targeted approaches offer better sensitivity, reduced
cost, and decreased complexity of bioinformatic analysis
15. CRISPR/Cas system:
an emerging
technology for
diagnostics
• Isothermal amplification technology,
relying on Cas protein, an
endonuclease that cleave
complementary sequences
• Cleavage induces nonspecific
cleavage of single stranded DNA or
RNA, which can be modified with
reporter/quencher, allowing signal
detection
• Applied to viral pathogen detection
Next-generation diagnostics with CRISPR | Science (sciencemag.org)
16. Future Diagnostics for Meningitis
Existing platforms for meningitis pathogens
• Most Category A pathogens
• High sensitivity and specificity
• Expensive
Platforms with the potential for Use Case 3
• Not developed for meningitis pathogens
• Meet many features outline in Use Case 3 TPP
• Lower cost
Advanced and emerging technologies for Use Case 3
• Sequencing or CRISPR/Cas based
• Early development for diagnostics
17. What’s next?
Assess various platforms-
existing/potential and
estimate market size
• Identify suitable platforms for
validation
• Develop and validate tests for
meningitis pathogens
• Estimate market size for
meningitis diagnostics
1
Pilot studies to inform
global deployment
strategies
• Evaluate RDT field performance
and lab capacity at local levels in
selected countries
• Assess LP rate, supply
chain/specimen transport
systems, and data reporting
2
Develop region/country-
specific rollout strategies
• Country’s risk level for
meningitis and prevalence of
pathogens
• Impact on surveillance, testing
algorithm, data flow etc
• Shift in the roles of laboratory at
national/subnational levels
• Procurement process/trainings
3
18. Conclusion
• RDTs are important for rapid meningitis detection at local
hospitals and laboratories; culture remains important for AMR
and genomic surveillance
• Strong partnership and innovations in technology and
informatics accelerate the development of next generation rapid
tests
• Deployment of new RDTs requires engagement of multi-
stakeholders and may lead to a paradigm shift in the roles of
clinical and public health laboratories
• Partner/country’s commitment and investment ensures
sustained access to RDTs
19. Acknowledgement
• Katya Fernandez and Olivier Ronveaux, WHO
• Use case 3 expert group (China CDC, FIND, PATH, IPP,
MSF, NICD S. Africa, NIPH, ULC, Uni. Of Liverpool, US
CDC, and WHO HQ)
• PATH landscape analysis team
• Meningococcal Working Group (GAVI, Bill and Melinda
Gates Foundation, UNICEF, WHO and CDC)
• CDC Meningitis and Vaccine Preventable Disease
Branch (lab/epi members)