1 / 23

Detection of Viruses by the Integrated Virus Detection System (IVDS)

Detection of Viruses by the Integrated Virus Detection System (IVDS). Dr. Charles Wick R&T Directorate U.S. Army Edgewood Chemical Biological Center Aberdeen Proving Ground, MD 21010 Charles.h.wick@us.army.mil; telephone: (410) 436-3321. Integrated Virus Detection System (IVDS). Outline.

nero
Download Presentation

Detection of Viruses by the Integrated Virus Detection System (IVDS)

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Detection of Viruses by the Integrated Virus Detection System (IVDS) Dr. Charles Wick R&T Directorate U.S. Army Edgewood Chemical Biological Center Aberdeen Proving Ground, MD 21010 Charles.h.wick@us.army.mil; telephone: (410) 436-3321

  2. Integrated Virus Detection System (IVDS) Outline • Introduction to IVDS • Results of other viruses • Honey Bee Results

  3. Rapid Virus Detection Integrated Virus Detection System • Concept • Instrument and Sample Data • Fate curves • Viruses in mixtures • Honey Bee Results • References

  4. Integrated Virus Detection System • Gathers wide ranged information about virus threats • Not limited by reagents • Not limited by prior knowledge • Detection of multiple viruses in one analysis. • Detects unknown and modified viruses • Detects empty virus capsules • Commercialized/COTS

  5. Integrated Virus Detection System Counting means Fast Virus Counting Platform Utilizes “Virus Window” Accumulate over time (additional advantage) Concentrate (10-4 mechanical Advantage) Virus Sample Automatic recording of virus concentration and size Counts intact virus particles without PCR, antibodies or reagents Detection of all viruses types – enables detection of all viruses, including unknowns. Multiple viruses counted in same sample in a few minutes.

  6. 1 2 3 Integrated Virus Detection System Sample Analysis Virus Size Distribution Table Graph of Concentration vs. Virus Size Detailed Data

  7. Integrated Virus Detection System MS2 stability vs temperature at 70oC 2001 ILIR Review: Temperature Fate Curve Percent intact for 70C: 90% at 1 minute 50% at 3 minutes 20% at 10 minutes

  8. Integrated Virus Detection System Kilham Rat Virus KRV Ultrafiltration 100K UF, Buffer washed Salt peak removed revealing Parvovirus at 22.5 nm Unexpected contamination viruses at 34.6, 49.6 and 71nm (confirmed by other means). Multiple viruses counted in single sample.

  9. Integrated Virus Detection System Mouse Hepatitis Virus A59 MHV-A59 Neat Coated Coronavirus at 73.7 nm Uncoated virus at 51.4 nm Delicate viruses detected directly from source material.

  10. IVDS Bee Test 2_2 Grind 2 g bees (~20 bees + 30 ml AA) Pre-filter Centrifuge 20,000 rpm/60 min (~40,000 x g) Decant and filter through 0.2 um hollow fiber cartridge UF 300K 26 ml reduced to 3 ml Avg. of 5 scans each

  11. IVDS Bee Test 3 Grind 2 g bees (~20 bees + 30 ml AA) Pre-filter Centrifuge 20,000 rpm/60 min (~40,000 x g) UF 300K 30 ml reduced to 4 ml Avg. of 5 scans each

  12. IVDS Bee Test 4 Grind 2 g bees (~20 bees + 30 ml AA) Pre-filter Centrifuge 20,000 rpm/60 min (~40,000 x g) UF 300K 30 ml reduced to 4.5 ml Avg. of 5 scans each

  13. IVDS Bee Test 5

  14. IVDS Bee Test 6 CA #1

  15. IVDS Bee Test 7

  16. IVDS Bee Test 8

  17. IVDS Bee Test 9

  18. IVDS Bee Test 10 CA #2

  19. Honey Bee Summary • Detected, Comments • Yard #1, Sample F #131.1, 38.5nm -Test 2 • Yard #1, Sample F #2 [31.1nm] -Test 3 • Yard #1, Sample F #3 31.1nm, 38.5nm - Test 4 • Aussie #1, 38.5nm -Test 5 • Location #1, CA; 3/14/07; PA-Key; CAF3B; 1 Frame Bee 25, 33.4nm -Test 6 • Aussie #2, 27.9nm, [37.2nm] - Test 7 • Aussie #3, 26.9nm, [37.2nm] - Test 8 • Aussie #4, 27.9nm, [37.2nm] - Test 9 • Location #2, CA; 3/18/07; Strong hive 3; 10-20 frame, 27.9nm - Test 10 [low concentration]

  20. Integrated Virus Detection System • Work to date indicates the following: • Detect viruses in 5 minutes* • Determines presence of Threat • Determines absence of Threat • Gives Concentration and Size • Can be used in combination with other technologies. * Depending on sample source and need for preparation.

  21. Integrated Virus Detection System References • C.H. Wick, I. Elashvili, M. F. Stanford, P.E. McCubbin, S. V. Deshpande, Mass Spectrometry and Integrated Virus Detection System Characterization of MS2 Bacteriophage. Toxicology Mechanisms and Methods, Accepted for publication, (2006). • Kuzmanovic, D. A., I. Elashivili, C. H. Wick, C. O'Connell, and S. Krueger..Structural Analysis and Quantification of RNA in Bacteriophage MS2-like Viruses in Solution by Small Angle X-ray Scattering. 2005, Journal of Applied Environmental and Microbiology, accepted 2006. • Kuzmanovic, D. A., I. Elashivili, C. H. Wick, C. O'Connell, and S. Krueger.The MS2 Coat Protein Shell is Likely Assembled Under Tension: a Novel Role ofr the MS2 Bacteriophage A Protein as Revealed by Small-angle Neutron Scattering, J. Molecular Biology, 355, 1095-1111, (2006). • Wick, C.H., P.E. McCubbin, R.E. Jabbour, S.V. Deshpande, Detecting Bacteria by Direct Counting of Structural Protein Units by IVDS and Mass Spectrometry. Toxicology Mechanisms and Methods, 16: 485-493, (2006). • Wick, C.H., Kuzmanovic, D.A, Effects of Sample Impurities on the Analysis of MS2 bacteriophage by Small Angle Neutron Scattering, ECBC TR-441, (2005). • Kuzmanovic, D.A., I. Elashvili, C. H. Wick, C. O’Connell and S. Krueger, Bacteriophage MS2: Molecular Weight and Spatial Distribution of the Protein and RNA Components by Small-Angle Neutron Scattering and Virus Counting, Structure, Vol. 11, 1339-1348, (2003).

  22. Integrated Virus Detection System • Patents • Wick, C.H., 6, 491, 872 Method and system for detecting and recording submicron sized particles • Wick, C.H., 6, 485, 686 Method and apparatus for counting submicron sized particles • Wick, C.H., 6, 238, 866 Detector for nucleic acid typing and methods for using the same. • Wick, C.H., 6, 051, 189 System and method for detection, identification and monitoring of submicron-sized particles • Others pending

  23. Point of ContactCharles WickPhone: 410-436-3321Fax: 410-436-6783

More Related