1. r Diagnostics SARS Coronavirus Detection Karen K. Y. Young, Ph.D. Roche Molecular Diagnostics Thomas Emrich, Ph.D. Roche Applied Sciences

2. r Diagnostics Critical Issues Logistical challenges –Nature of epidemic not well understood Seasonal? Reservoir? –For rapid introduction of diagnostic assay Need to mobilize resources quickly Need partnership with regulatory agencies –Sample acquisition –IP landscape

3. r Diagnostics Critical Issues Technical challenges –Optimal sample type? Blood/serum/plasma Respiratory specimen Stool/urine –Sample preparation issues Removal of inhibitors Efficient lysis/preservation of RNA –Viral sequence heterogeneity Mutation Recombination –Standardization/reference material

4. r Diagnostics Roche’s Response Rapid development of research assay –Quantitative real-time PCR –LightCycler –Sample preparation defined by user Development of IVD assay if warranted –Regulatory guidance needed for performance requirements –Lessons learned from research assay –Automated sample preparation –TaqMan PCR –Full-process controls

5. r Diagnostics Research Assay Status LightCycler-based assay –Real-time PCR –Quantitative, external calibration standards –Internal control User-defined sample preparation In-house evaluation of positive samples completed World-wide trial at beta sites on-going

6. r Diagnostics Research Assay Performance In-house evaluation –Positive samples from Hong Kong (N=40) –Sample types Nasal-pharyngeal aspirates Stool Urine Throat swabs

7. r Diagnostics Research Assay Performance In-house evaluation

8. r Diagnostics Challenges to SARS diagnosis Many unknowns –Nature of epidemic? –Natural history of infection? –Most appropriate sample type? –Optimal sample handling?

9. r Diagnostics Challenges to SARS diagnosis To effect rapid introduction of diagnostic assay –Requires dedicated resources –Co-operation from regulatory/public health agencies needed Definition of performance requirements Standardization for assessment of assay performance Availability of reference standards Availability of clinical samples for assay development and evaluation