1. Real-Time RT-PCR Influenza Virus Detection from Blood Samples Collected in PrimeStore MTM 1747 Citadel Plaza Suite 206 San Antonio, TX 78209 (210) 826-0910 ltd@lhnvd.com Luke T. Daum, Ph.D.

2. Methods into Inactivation Inactivation, RNA stabilization Real-time RT-PCR detection 1.5 mL PrimeStore MTM 0.5, 0.3 or 0.1 mL of blood spiked with 3 concentrations of FluA virus Extraction ABI 7500

3. Is blood collection compatible with PrimeStore MTM and does blood concentration affect downstream real-time RT-PCR detection? SPECIMEN COLLECTION & PRESERVATION SOLUTION FOR MOLECULAR DIAGNOSTIC APPLICATIONS PrimeStore® 0.5, 0.3 and 0.1 mL of blood matrix + Flu A was inoculated into standard 1.5 mL tubes of PrimeStore

4. Influenza A Detection from varying blood concentrations

5. Answer 10^5, 10^4 and 10^3 TCID50/mL of Influenza virus were readily detected from blood matrices collected in PrimeStore MTM Cycle thresholds (CTs) obtained by real-time RT-PCR using an ABI 7500 showed little variation using 0.5, 0.3, or 0.1 mL blood in PrimeStore MTM tubes.

6. Does detection of virus from blood collected in PrimeStore MTM improve extraction efficiency compared to equilavant amounts of virus in water (control) or a commerically available lysis solution? SPECIMEN COLLECTION & PRESERVATION SOLUTION FOR MOLECULAR DIAGNOSTIC APPLICATIONS PrimeStore® 0.5 mL of blood in PrimeStore, nuclease- free water, or commercial lysis solution

7. Virus Detection from Blood in PrimeStore MTM Cycle Threshold Note: Lower real-time CT value = higher initial viral RNA targets

8. Answer Sensitivity of influenza A virus was enhanced prior to extraction by collection in PrimeStore MTM compared to nuclease free water or a standard commercial lysis buffer. Improved CT was for initial (Day 0) extraction. The preservation and stability in PrimeStore continues over time (data not shown) with degradation noted in water and commercial lysis buffer.

9. Longhorn Vaccines & Diagnostics LLC 1747 Citadel Plaza, Suite 206 San Antonio, Texas 78209 Phone: (210) 826-0910 Email: info@lhnvd.com Luke T. Daum, Ph.D. QUESTIONS?