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Total Bioburden Assays 04 06 2010
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Total Bioburden Assays 04 06 2010

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IMT\'s Total Bioburden Assays (TBA) represent the cutting-edge of Total Bacteria Count (TBC) and Total Yeasts and Molds Count (TYC) tools.

IMT\'s Total Bioburden Assays (TBA) represent the cutting-edge of Total Bacteria Count (TBC) and Total Yeasts and Molds Count (TYC) tools.

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  • 1. IMT’s Total Bioburden Assays IMT’ T t l Bi b d A Andy Moreno d Vice President, Microbiology Sales Engineering Cell: +559 827 8245 Cell: +559‐827‐8245 FAX: +866‐776‐3746 andy.moreno@imt‐pcr.com http://www.imt‐pcr.com Linkedin Profile Skype: Andy Moreno Sk pe And Moreno Slide 1 of 13
  • 2. Industrial Microbial Testing – Based in Newport Beach, Calif., manufactures and  markets assays to support rapid real‐time  quantitative PCR microbial testing.  – IMT's One‐Step Total Bacteria Count (TBC)  and  Total Yeasts and Molds Count (YMC) T t lY t d M ld C t (YMC) quantitative  tit ti real‐time polymerase chain reaction (qRT‐PCR)  assays replace standard plate count methods. assays replace standard plate count methods Slide 2 of 13
  • 3. TRADITIONAL PLATE COUNT (48 HOURS) Slide 3 of 13
  • 4. TOTAL BACTERIA COUNT BY PCR (1 HOUR) DNA dilutions after boil prep  for PCR Real‐Time  Standard  Data Curve C Slide 4 of 13
  • 5. Slide 5 of 13
  • 6. “Organism Prospecting Package” • Water/matrix/Ingredients/In Process/Final  Product sample • Total Bacterial Count (TBC) screen •Is there bacteria?  •Presence/Absence Quantitation •Quantitation •Gram Negative/Gram Positive •Total Yeasts and Molds Count (YMC) screen •Are there yeasts and molds?  •Presence/Absence •Quantitation Slide 6 of 13
  • 7. IMT’s TBA: “Internal Investigation Tool” • Any matrix, post sample preparation • 30‐minute run time in SmartCycler 30 i t ti i S tC l • Sensitivity to one (1) cell for TBC • Presence/Absence P /Ab • Quantification Slide 7 of 13
  • 8. IMT’s OneStep Assays ~30 minute run time 30 minute run time 85 µl molecular‐ 5 µl purified DNA  25 µl Total reaction  grade water added  grade water added Sample volume in  volume in to primers & probes  SmartCylcer tube included in dried‐ down reagents Slide 8 of 13
  • 9. Typical QA/QC Protocol • IMT TBA Assays and Cepheid SmartCycler IMT TBA Assays and Cepheid SmartCycler • Minimally‐trained QA Team • 30 minute easy to use cheap results 30 minute, easy‐to‐use, cheap results Slide 9 of 13
  • 10. Collaborative Study to evaluate a Total Bacteria Count  Assay using Q A i Quantitative Real‐Time PCR tit ti R l Ti PCR A collaborative study was undertaken to demonstrate the precision and  accuracy of a total bacteria count (TBC) assay developed to run with  accuracy of a total bacteria count (TBC) assay developed to run with commercially available PCR reagents and a rapid thermocycling platform.  The TBC assay can be completed within one hour.  The TBC  assay has many applications for enhanced HACCP applications  The overall conclusions are that all the labs performed well in  constructing the standard curve and had some variance with the  unknown samples.  However, the results show that all collaborators  successfully used the TBC assay to quantify bacterial load accurately  f ll d h C if b i ll d l within one hour.  The TBC assay can be used for a rapid real‐time  measure of bacteria in many diverse applications and support HACCP  for real‐time statistical process control.  Slide 10 of 13
  • 11. IMT Testing Assays ‐Screen for presence/absence, quantitation ‐Total Bacteria Count (TBC) ‐Total Yeasts and Molds Count (YMC) ‐Salmonella ‐E. coli species E li i ‐O15:H7 ‐Listeria ‐C. botulisum “A”, “B”, “C” and “E” Slide 11 of 13
  • 12. Quantitative Real Time PCR Systems Quantitative Real‐Time PCR Systems Eppendorf Slide 12 of 13
  • 13. Questions & Answers Questions & Answers Slide 13 of 13