Testing Methodologies and Sampling

[Transcript of Dr. Tortorello's remarks]

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Authors: Mary Lou Tortorello, Ph.D. and Tong-Jen Fu, Ph.D., National Center for Food Safety and Technology

**Table of Contents**
Testing Methodologies and Sampling Overview Why test? Pre-guidance questions on microbiological testing What should be tested? Difference in microbial counts between sprouts and water Detection of Salmonella in Sprouts and Water from Outbreak Seeds Pre-guidance questions on microbiological testing How to test: requirements for guidance testing methods Testing of Water From each Production Batch for: Salmonella and E-Coli O157:H7 Testing of Water From each Production Batch for Detection of Pathogens In Inoculated Sprouts and Sprout Water by Rapid Test Kits Detection Limit of Rapid Test Kits Without Enrichment Growth of Salmonella During Sprouting of Outbreak Seeds Detection of Salmonella in Sprouts and Water from Outbreaks Seeds Microbiological testing: validation of rapid tests Validation of Salmonella Detection by Rapid Tests Assurance Gold and VIP Selected AOAC Performance Tested Methods Detection Limit of Rapid Test Kits for Salmonella in Buffer Detection of Salmonella in Sprouts and Water from Outbreak Seeds by Rapid Test Kits and BAM Publications on Pathogen Detection Methods in Sprouts Method Considerations Methodologies and Sampling.Looking Forward Sampling and Testing: New Issues Sampling and Microbial Testing of Spent Irrigation Water When to Sample: 24 h vs. 48 h	Growth of pathogens during sprouting of alfalfa seeds Summery of Literature Data pathogen growth at 24 h and 48 h of sprouting How to Sample Sample Pre-Concentration Standard Membrane Filtration Tangential Flow Filtration System Evaluation of TFF for Concentration of Pathogens Validation of Tangential flow filtration as part of sample treatment for detection of Salmonella in spent sprout irrigation water Recovery of Salmonella from the TFF system Validation of TFF System: E. coli 0157:H7 Recovery of E. coli 0157:H7 from the TFF system Seed Sampling and Testing Isolation of pathogens from outbreak associated sprouts/seeds Detection of Salmonella in Naturally Contaminated Seeds(Inami et al., 2001) What to Test: Seeds vs. Sprouts.Inami et al., 2001 What to Test: Seeds vs. Sprouts.Fu et al., 2005 (unpublished) Effect of Sampling Size Screening Seeds Used for Sprout Production: Industry Practice Detection Methodologies Automated Immunomagnetic Separation System Evaluation of the Pathatrix IMS system for isolation of E. coli 0157:H7 from spent irrigation water Evaluation of the Pathatrix IMS system for isolation of Salmonella from spent irrigation water Real Time Pathogen Monitoring Fiber-Optic Biosensor Literature Data Evaluation of the Analyte 2000? Fiber-Optic Biosensor for Detection of E. coli O157:H7 in Spent Sprout Irrigation Water Conclusions Acknowledgements