VaporPrintTM of Infectious Bacteria Cultures Strep pneumonia Shigella flexneri Salmonella typhimurium Klebsiella pneumonia Pseudomonas aeruginosa Candida albicans Fresh Nutrient Staph aureus Hemophilus influenza Electronic Sensor Technology Salmonella enteritidis Enterococcus faecalis E. Coli E. Coli O157 Cultures Prepared by: Department of Pathology and Laboratory Medicine, State Public Health Laboratory of Nevada WWW.ESTCAL.COM 1 An Investigation of Infectious Bacteria With a GC/SAW Electronic Nose • • • • • Water based cultures were inoculated with 12 infectious bacteria and allowed to incubate at 37oC for 24 hours before analyzing headspace vapors with GC/SAW Electronic nose. Two of the twelve cultures did not develop and contained no bacteria. Headspace vapor of cultures without bacteria displayed two primary nutrient analyte peaks and three small analyte peaks located midway between the two nutrient peaks. Bacteria growth dynamically interacted with nutrient compounds as well as created new analyte vapors specific to the bacteria itself. The concentrations of new analytes together with modified nutrient vapors created VaporPrintsTM images specific to bacteria type. E.Coli and E.Coli )157 produced nearly identical VaporPrintsTM, however, one analyte was found to be specific to only O157. In general each bacteria type could be recognized by its unique VaporPrintTM image. Bacteria Tested 1. Strep pneumonia 2. Hemophilus influenza 3. Candida albicans 4. Shegella flexneri 5. Staphylococcus aureus 6. Pseudomonas aeruginosa 7. Salmonella enteritidis 8. Enterococcus faecalis 9. Klebsiella pneumonia 10 Escherichia coli O157 11 Escherichia coli (benign) 12 Salmonella typhimurium Electronic Sensor Technology Ease of Summary of Results Identification 0 0 4 5 3 10 7 5 10 10 10 10 Negative Culture Growth Negative Culture Growth Doubled 2nd nutrient peak and 2 persursors Nutrients diminished, percursor and precursor ripples Increased nutrients, doubled 2nd Reduced 1st nutrient, 10X new ripple peak Reduced 1st nutrient, Large percursor peak Reduced 1st nutrient, Large increase in 2nd Reduced nutrients, large new ripple and percursors Reduced 1st, increased 2nd, large post, added ripple Same as O157 except 4th ripple peak not present Reduced nuturients, large new ripple, precursors WWW.ESTCAL.COM 2 No Growth Confirmed for Hemophilus influenza and Strep pneumonia Culture Bottles Strep pneumonia Nutrient Broth Hemophilus influenza Nutrient Broth Analytes Present in Nutrient Broth Primary Nutrient Peaks (2) Ripple Peaks (3) Post Peaks (1) Electronic Sensor Technology Retentio n Time Amount after 24 hr @ 37 o C 6.8 10.3 7.8 8.2 9 12.5 362 749 10 26 151 55 Quantitative results based upon no-growth chromatograms of H. influenza and S. pneumonia Small set of 3 peaks prominent and generally always present but much smaller than primary nutrient peaks Broad low vapor pressure compounds generally present WWW.ESTCAL.COM 3 Candida albicans Analytes Present in Candida albicans Primary Nutrient Peaks (2) Ripple Peaks (3) New Peaks (2) Post Peaks (1) Retentio n Time Amount after 24 hr @ 37 o C 6.8 10.2 7.8 8.2 9.0 3.14 5.26 12.26 344 1383 10 26 169 40 111 127 First peak unchanged but second doubled ripple peaks unchanged Two new perecursor peaks, very distinctive Doubled heavy compounds Candida albicans Nutrient Broth Electronic Sensor Technology WWW.ESTCAL.COM 4 Shigella flexneri Analytes Present in Shegella flexneri Primary Nutrient Peaks (2) Ripple Peaks (3) Precursor Peaks (4) Post Peaks (1) Retention Time Amount after 24 hr @ 37 o C 7.1 10.5 7.8 8.2 9.0 3.22 4.9 5.42 5.98 12.26 172 399 10 26 155 30 26 23 11 32 First peak almost gone and second greatly reduced ripple peaks unchanged (1) percursor peak (3) new persursor ripple peaks unchanged small mixture Shigella flexneri Nutrient Broth Electronic Sensor Technology WWW.ESTCAL.COM 5 Staphylococcus aureus Analytes Present in Staphylococcus aureus Primary Nutrient Peaks (2) Ripple Peaks (3) Post Peaks (1) Retention Time Amount after 24 hr @ 37 o C 6.74 10.22 7.8 8.2 9.0 12.26 522 1711 10 26 155 159 First peak slight increase and second doubled++ ripple peaks unchanged well defined unchanged Staph aureus Nutrient Broth Electronic Sensor Technology WWW.ESTCAL.COM 6 Pseudomonas aeruginosa Analytes Present in Pseudomonas aeruginosa Primary Nutrient Peaks (2) Ripple Peaks (3) Precursor Peak (1) Post Peak (1) Retention Time Amount after 24 hr @ 37 o C 6.78 10.22 7.8 8.2 8.9 3.1 12.26 202 330 10 26 5170 31 32 First peak almost gone and second slightly reduced first two ripple peaks unchanged but third is 10 times larger than nutrient peaks (1) percursor peak unchanged small mixture Pseudomonas aeruginosa Nutrient Broth Electronic Sensor Technology WWW.ESTCAL.COM 7 Salmonella enteritidis Analytes Present in Salmonella enteritidis Primary Nutrient Peaks (2) Ripple Peaks (3) Precursor Peak (1) Post Peaks (1) Retention Time Amount after 24 hr @ 37 o C 6.94 10.18 7.8 8.2 9.0 3.1 11.5 250 546 10 28 155 327 25 First peak almost gone and second unchanged ripple peaks unchanged (1) percursor peak as big as nutrient peaks unchanged small mixture Salmonella enteritidis Nutrient Broth Electronic Sensor Technology WWW.ESTCAL.COM 8 Enterococcus faecalis Analytes Present in Enterococcus faecalis Primary Nutrient Peaks (2) Ripple Peaks (3) Precursor Peak (1) Post Peaks (1) Retention Time Amount after 24 hr @ 37 o C 6.74 10.18 7.8 8.2 9.0 2.34 12.26 391 1693 10 26 155 16 141 First peak slight decrease and second greatly increased ripple peaks unchanged (1) small percursor peak distinctive small peak Enterococcus faecalis Nutrient Broth Electronic Sensor Technology WWW.ESTCAL.COM 9 Klebsiella pneumonia Analytes Present in Klebsiella pneumonia Primary Nutrient Peaks (2) Ripple Peaks (4) Precursor Peaks (2) Post Peaks (1) Retention Time Amount after 24 hr @ 37 o C 6.94 10.26 7.8 8.2 9.0 9.5 1.02 3.1 11.46 272 169 10 26 155 549 30 165 169 Both nutrient peaks reduced original (3) ripple peaks unchanged but with a fourth large new peak (2) distinctive percursor peaks large grouping of post peaks Klebsiella pneumonia Nutrient Broth Electronic Sensor Technology WWW.ESTCAL.COM 10 Escherichia coli O157 Analytes Present in Escherichia coli O157 Primary Nutrient Peaks (2) Ripple Peaks (4) Post Peaks (1) Retention Time Amount after 24 hr @ 37 o C 6.7 10.26 7.8 8.2 9.0 9.5 11.22 129 510 10 26 155 100 2135 early peak reduced and second peak increased original (3) ripple peaks unchanged but with a fourth new peak large new post peak E. coli O157 Nutrient Broth Electronic Sensor Technology WWW.ESTCAL.COM 11 E. coli (benign) E. Coli (benign) Nutrient Broth Analytes Present in Escherichia coli (benign) Primary Nutrient Peaks (2) Ripple Peaks (3) Post Peaks (1) Electronic Sensor Technology Retention Time Amount after 24 hr @ 37 o C 6.7 10.26 7.8 8.2 9.0 11.22 129 510 10 26 155 4002 early peak reduced and second peak increased original (3) ripple peaks unchanged large new post peak WWW.ESTCAL.COM 12 Comparison of E. coli E. coli O157 E. coli (benign) Specific to O157 E. Coli Electronic Sensor Technology WWW.ESTCAL.COM 13 Salmonella typhimurium Salmonella typhimurium Nutrient Broth Analytes Present in Salmonella typhimurium Primary Nutrient Peaks (2) Ripple Peaks (3) Precursor Peak (2) Post Peaks (1) Electronic Sensor Technology Retention Time Amount after 24 hr @ 37 o C 6.86 9.94 7.8 8.18 9.0 0.94 3.42 10.9 608 522 10 1792 155 1281 175 25 WWW.ESTCAL.COM First peak reduced and second almost gone large new ripple peak (2) distinctive percursor peaks many low volaltility analytes present 14