im ts food qa qc assays 07 07 2010

IMT’s Food QA/QC Assays

Andy MorenoVice President, Microbiology Sales Engineering

Cell: +559-827-8245FAX: +866-776-3746

andy.moreno@imt-pcr.comhttp://www.imt-pcr.com

Linkedin ProfileSKYPE: Andy Moreno USA

Slide 1 of 8

Industrial Microbial Testing (IMT)

Manufactures assays to support rapid real-time quantitative Polymerase Chain Reaction microbial testing. • Actionable Information: Rapid Release/99% product safe• Establish Positive Release Program• Food Safety Screening: Hold/Release time

– ~One (1) hour/PCR vs. Traditional Culture Media Testing – Ingredients, In-process Solutions, Finished Product

• Sensitivity: One (1) cell per 5 µl of sample• One (1) sample, many tests

Slide 2 of 8

Real-Time Data

Standard Curve

Traditional Culture Media vs. ~1 HOUR qRT-PCR

versus3 to 14 days ~ One (1) Hour

Slide 3 of 8

Slide 4 of 8

“Organism Prospecting Screens”

Water or Matrix• Irrigation water/Ingredients/In Process/Final Product sample

• Total Bacterial Count (TBC) screen•Is there bacteria?

•Presence/Absence

•Quantitation

•Total Yeasts and Molds Count (YMC) screen•Are there yeasts and molds?

•Presence/Absence

•Quantitation

Slide 5 of 8

IMT’s OneStep Assays

85 µl molecular grade water added to primers with PCR chemistry in dried-down format

5 µl purified DNA Sample per Cepheid Tube plus 20µl water and primer chemistries: total 25 µl—4 test batch

~30 minute run time

Slide 6 of 8

Typical Food QA Station & Team

• IMT TBC Assay and Cepheid SmartCycler• Minimally-trained QA/QC team(s)• 30 minute, easy-to-use, accurate DNA results

Slide 5 of 7

IMT Assays• Total Bioburden Assays

– OneStep Total Bacteria Count Assay– OneStep Total Yeasts & Molds Count Assay

• Commercialized Pathogen Assays– E. coli spp. Assay– Salmonella spp. Assay– C. botulinum “E” Assay– Porcine Assay

• R&D Pipeline– O157:H7– Campylobacter Assay– Listeria mono. Assay– Enterococcus spp. Assay– Staphylococcus spp. Assay– Pseudomonas spp. Assay– Xanthomonas spp. Assay– Mycoplasma spp. Assay

Slide 8 of 8

Samples

TotalBacteria

Count

Pathogenof

Interest

PositiveReleaseProgram

IMT’s Assays Validate Food Producer’s Statistical Process Control Program

Periodic Testing of Randomized samples against established maximum and minimum standards.

Food: Positive Release Program

•Irrigation water tested

•Raw fruit and vegetables are washed and cleaned

•Peracetic Acid and Chlorine wash bath tested at the end of each

run

•Finished product is tested

•Environmental samples collected in plant “hot spots”

Slide 10 of 8

Sample Environmental Testing*

Surface Sample Collection Collect bacterial samples in filter One sample, 5 tests-Total Bacterial Count-E. coli spp.-Salmonella spp.-Listeria spp.

Collection and preparation of samples = 30 minutes Quantitative real-time PCR results in 30

minutes

*Quantitatively track the presence pathogens in the environment and correlate with spoilage using statistical process control methodologies.