A Field-Deployable Insulated Isothermal PCR-Based System for Rapid and Sensitive Detection of Avian Influenza a Virus at Points of Need
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POCKIT Central PCR System for Detecting African Swine Fever Virus in Vietnam The document describes a study evaluating the POCKIT Central PCR system for detecting African Swine Fever Virus (ASFV) in Vietnam. The system provides fully automated sample-to-answer detection of ASFV in 85 minutes using cartridges that integrate nucleic acid extraction and PCR. The study found the POCKIT Central system performed equivalently to the OIE reference real-time PCR method, with high analytical sensitivity and specificity. The automated system reduces hands-on time and human error compared to traditional PCR methods. It provides a simple workflow for rapid on-site detection of ASFV to help control the ongoing spread of the disease
A Field-Deployable Insulated Isothermal PCR-Based System for Rapid and Sensitive Detection of Avian Influenza a Virus at Points of Need
- 1. GeneReach GeneReach A Fully Automated Sample-to-Answer POCKIT™ Central PCR System for Detecting African Swine Fever Virus in Vietnam Simon Chung GeneReach
- 2. GeneReach African Swine Fever is Spreading in Asia Can it be controlled? Can it be controlled? Can it be controlled? 2
- 3. GeneReach Government Farm 1 Report Suspect Case Investigation Dx and Confirmation Farm 1 Farm 2 Farm 3 On Fire Put on the fire Current Control Strategy 3
- 4. GeneReach Put Out the Fire vs Current ASF Control Strategy Put out the fire ASF prevention and control by farmers • Disinfection • Biosecurity • Close the farm for self quarantine • Who is the next? Afraid 4
- 5. GeneReach Background Biosecurity check Pulling the bad teeth out • Spot stamping out Pig producers are using PCR to detect the ASFV for prevention and control of ASF in endemic area The PCR system they are using should be validated 5
- 6. GeneReach Objective Analytical sensitivity Analytical specificity Diagnostic performance vs. oie real time PCR method 6 POCKIT™ Central ASFV detection system
- 7. GeneReach Materials and Methods Samples and locations POCKIT Central vs real time PCR Workflow 7
- 8. GeneReach Samples and Locations • Lymph nodes • Tonsil • Spleen • Serum National Center of Veterinary Diagnosis Ha Noi Regional Animal Health Office 6 Ho Chi Minh Tissue samples are homogenized by tacoPrep bead beating homogenizer 8
- 9. GeneReach POCKIT™ Central vs oie Real Time PCR Integrates NAs extraction and PCR King et al. 2013 Interpretation and cut off NCVD: Ct < 35, ASFV positive No Ct detected, ASFV negative 35 ≤ Ct ≤ 45, suspicious sample RAHO6: Ct < 40, ASFV positive No Ct detected, ASFV negative 40 ≤ Ct ≤ 45, suspicious sample Results Description + Positive, ASFV nucleic acid detected - Negative, ASFV nucleic acid not detected ? Undetermined, repeat reaction from original sample ! Waring ＊ Abnormal, signals outsides detection range Extraction system NCVD: taco 24 taco DNA/RNA Extraction Kit RAHO6: Thermo Scientific KingFisher Purification system 9
- 10. GeneReach Workflow Sample preparation NAs extraction PCR Interpretation Sample preparation POCKIT Central Analytical sensitivity Analytical specificity Comparison of diagnostic performance Analytical sensitivity Analytical specificity Comparison of diagnostic performance Real Time PCR 85 minutes 50 minutes 80 minutes 10
- 11. GeneReach POCKIT™ Central Operation Procedure Experiment setup 11
- 12. GeneReach Results Analytical sensitivity Analytical specificity Comparison of diagnostic performance 12
- 15. GeneReach Comparison of Diagnostic Performance NCVD RAHO 6 15
- 16. GeneReach Conclusions • Performance of POCKIT Central ASFV Reagent is equivalent to oie reference real time PCR method in NCVD and RAHO 6 Dx labs. • The sample type can be tissue homogenate and serum from the pig. • The turn around time of POCKIT Central is shorter than real time PCR method. • The cartridge of NAs extraction and consumables of POCKIT Central simplifies the preparation of traditional PCR procedures. • The lyophilized PCR premix of POCKIT Central ASFV reagent simplifies the PCR preparation time and reduces the human error. 16
- 17. GeneReach After ASF spread in Asia… Biosecurity Cleaning Disinfection Detection Check the pigs by PCR Pulling bad teeth Biosecurity Keep production 17
- 18. GeneReach Take Home Messages • Equivalent to oie real time PCR method Good Performance • Fully automated • Sample in and answer out Reduce human error • Simple and universal workflow Farm side detection • Turn around time is 85 minutes Instant decision making 18
- 19. GeneReach NCVD RAHO 6 Vietchem R & D Team of GeneReach Technical support team of GeneReach Acknowledgement 19 Dr. David Tsai Mr. Pin Hsing Chou Ms. Yu Chun Lin Ms. Mark Tsai Ms. Nina Chen Ms. Luu Van Quynh Ms. Dam Thi Vui
- 20. GeneReach GeneReach Thank You for the Attention