One Health: Nano-Assembly Based Biosensor for Rapid Detection of Infectious Diseases
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GRF 2nd One Health Summit 2013: Presentation by Evangelyn C. Alocilja, Michigan State University
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One Health: Nano-Assembly Based Biosensor for Rapid Detection of Infectious Diseases
- 1. One Health: Nano-Assembly Based Biosensor for Rapid Detection of Infectious Diseases Evangelyn C. Alocilja, Ph.D., Professor Michigan State University 2nd One Health Summit 2013 17-20 November 2013, Davos, Switzerland
- 2. Main Idea R – rapid I – inexpensive S – simple E – early Detection Early Treatment Early Recovery Lab based Productive Life POC, Personal Sustainable Planet
- 3. Outline • Comorbidity diagnostic framework – to quickly diagnose comorbid health cases • Rapid disease detection involving humananimal interfaces • M. tuberculosis vs M. bovis • E. coli O157:H7 • Biosensor design, assay, and results • Conclusion
- 4. Comorbidity Diagnostic Framework Social Behavior Medical infra Religious Human Food Cultural Water/Air Finance Nano-enabled Policy diagnostic Chemicals systems for comorbid cases Technologies Economics Animal Institutional Infectious agents Globalization Security Ecological
- 5. Diagnostic Mega-Challenges Immunecompromised, elderly, very young Emerging infectious diseases Medical infrastructure Drug resistance: MDR, XDR, TDR
- 6. Example: TB/HIV Coinfection TB Infected 2.3 B New cases 7.6 M HIV 34 M 2.5 M TB/HIV Deaths 1.3 M 0.4 M 1.0 M 1.1 M Each facilitates the progression of the other. Drug-resistant TB Zoonotic TB “We cannot win the battle against AIDS if we do not also fight TB. TB is too often a death sentence for people with AIDS. …” Nelson Mandela July 15, 2004, "Confronting the Joint HIV/TB Epidemics", XV International AIDS Conference, Bangkok, 2004. http://www.tbfacts.org/tb-hiv.html http://www.who.int/hiv/data/en/ http://www.tbfacts.org/tb-hiv.html
- 7. Example: TB/Diabetes Infected New cases TB 2.3 B 7.6 M Diabetes 382 M TB/Diabetes 10% of global TB Deaths 1.0 M 5.1 M Higher risk of death during treatment People with diabetes have a 2-3 times higher risk of TB compared to people without diabetes. People with TB should be screened for diabetes and people with diabetes should be screened for TB. http://www.tbfacts.org/tb-hiv.html http://www.who.int/tb/publications/diabetes_tb.pdf
- 9. M. Tuberculosis vs M. bovis M. tuberculosis M. bovis Genomics Morphology Symptoms Treatment 99.95% similar similar similar isoniazid, rifampin, pyrazinamide, and ethambutol HIV Current diagnostic High incidence Do not distinguish the two Specific test Resistant to pyrazinamide Laborious, expensive, time consuming
- 10. M. Tuberculosis vs M. bovis M. Tuberculosis cell envelope Lipoprotein M (LprM) is deleted in M. bovis M. tuberculosis ESAT-6 + LprM + http://www.nature.com.proxy1.cl.msu.edu/nature/j ournal/v406/n6797/fig_tab/406788a0_F2.html M. bovis + http://www.nature.com.proxy1.cl.msu.edu/nm/j ournal/v13/n3/fig_tab/nm0307-286_F1.html -
- 11. Biosensor Schematic Extraction Detection Signal amplification Prototype 1 Next gen: Cell phone enabled RISE – Rapid, Inexpensive, Sensitive, Early
- 12. Bio-inspired Design Nano-structure: Peacock Design Head Body Tail-eyes Figure 1 ESAT-6 MNP-Ab Ab-Au-CdS lprM MNP-Ab Protein-based detection. Ab-Au-PbS
- 13. Nano-Assembly: Peacock Design Head Body Tail-eyes MNP-probe1 probe2-Au-CdS MNP-probe1 DNA-based detection. probe2-Au-PbS
- 14. Head: Magnetic Nanoparticles (MNPs) Oxidizing agent + Fe2O3 core + + EAM + + + + EAM + + + HCl EAM Aniline + - EAM - EAM Monoclonal antibody Antibody Modified EAM EAM EAM + DNA Probe EAM Labeled DNA Probe
- 15. Peacock Tail: AuNPs and NTs Gold NP solution during synthesis. Left to right: ZnS, CdS, and PbS. nanoparticle tracers in solution. TEM image of CdS, ~10nm. TEM images of AuNPs, ~15 nm in diameter. Welldispersed, uniform size and shape. TEM image of PbS, ~3nm. Oligonucleotide sequence: 5’-GTC AGT CAG TCA GTC AGT CA-3’ 3’ thiol modifier and 5’ amino modifier
- 16. Assay and Detection Wash Buffer Magnet Waste outlet Two-antibody system increases specificity
- 17. E. coli O157:H7 Abm - LPS & O antigen; Abp - O & H antigen cell http://www.ecl-lab.com/en/ecoli/index.asp
- 18. Capture Efficiency (%) - E. coli O157:H7 cell 100 C a p t u r e E f f i , c i % e n c y 90 80 70 60 50 40 30 20 10 0 10^1 10^2 1 2 PBS 10^3 3 10^1 10^2 10^3 10^1 10^2 10^3 10^1 10^2 10^3 4 5 6 7 8 9 10 11 12 Spinach Milk Apple Juice Axis Title Bacterial concentration, cfu/mL Specificity evaluation: 39 E. coli O157:H7 related and unrelated strains
- 19. E. coli O157:H7 Detection cell SWAV sensorgram of Au-PbS tail for E. coli O157:H7 detection at 103 cfu/ml.
- 20. Summary cell Concentration (cfu/ml) Average Current Peak (µA) Standard Deviation (µA) S/N Blank Avg. + 3*S.D. P value 0 101 102 103 104 105 106 4.33 10.25 12.94 13.33 13.75 14.83 18.67 1.36 4.50 1.77 4.08 2.87 4.78 1.00 2.37 2.99 3.08 3.17 3.42 4.31 0.003 0.000 0.000 0.001 0.011 0.000 N/A 8.42 N/A
- 21. Multiplex DNA-Based Assembly 0.00007 Control 500 pg/mL 5 ng/mL 50 ng/mL 0.00006 0.00005 Current (A) 50 pg/mL 0.00004 0.00003 0.00002 0.00001 0 -1.5 -1 -0.5 Potential (V vs. Ag/AgCl) 0
- 22. B. anthracis and Salmonella Enteritidis 50 45 40 y = 2.1498Ln(x) + 47.99 R2 = 0.9709 y = 2.1498Ln(x) + 44.99 R2 = 0.9865 Bacillus anthracis Salmonella Enteritidis 0.00001 Log. (Bacillus anthracis) Log. (Salmonella 0.0001 0.001 0.01 0.1 Enteritidis) Target DNA Concentration (ug/mL) Current (uA) 35 30 25 20 15 10 5 0 1
- 23. Summary • • • • • • • • Rapid: < 1 hr Inexpensive: $2/test Sensitive: 101-106 cfu/ml Extra-reliable: SNR = 2.37-4.31 Simple, handheld Multiplex detection Field operable Environmentally responsive cell Useful to both human and animal services
- 24. Value of Biosensor • Tool to differentiate source of infection • Understand human-to-human as well as humanto-animal transmission routes • Onsite diagnosis will be cost effective • Allows quick dispensation of treatment • Facilitates control of infection • Supports the implementation of One Health approaches. • Tool that could open up new frontiers for disease elimination. • Easily adapted to address many other diseases of global importance.
- 25. Acknowledgment of Funding Sources
- 26. Acknowledgment of Research Group • • • • • • Graduate students Postdocs Professorial assistants High school interns Co-investigators Collaborators For a complete listing, refer to: http://www.egr.msu.edu/~alocilja Thank you for your attention !
Notes de l'éditeur
- 25 min presentationL&G, good morning. Thank organizer for invitation; thank audience for coming
- Comorbidity – a concomitant but unrelated pathological or disease process. (The American Heritage® Medical Dictionary Copyright © 2007)
- COPD -chronic obstructive pulmonary diseaseEncopresis: The inability to control the elimination of stool.
- Ionic interaction between Pani and Ab; orients the Ab properly due to the – charge of the Fc region
- The biosensor has great potential as a tool in detecting trace quantities of mycobacterial antigens. For a multiplex system, the biosensor can be used to differentiate source of infection, understand human-to-human as well as human-to-animal transmission routes. Onsite diagnosis will be cost effective, will allow quick dispensation of treatment, will facilitate control of infection, and will support the implementation of One Health approaches.Biosensor will be an added tool for both human and animal health services.