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

GeneReach
GeneReach
A Fully Automated Sample-to-Answer
POCKIT™ Central PCR System for Detecting
African Swine Fever Virus in Vietnam
Simon Chung
GeneReach

GeneReach
African Swine Fever is Spreading in Asia
Can it be controlled?
Can it be controlled?
Can it be controlled?
2

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

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

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

GeneReach
Objective
Analytical sensitivity
Analytical specificity
Diagnostic performance
vs.
oie real time PCR method
6
POCKIT™ Central ASFV detection system

GeneReach
Materials and Methods
Samples and locations
POCKIT Central vs real time PCR
Workflow
7

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

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

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

GeneReach
POCKIT™ Central Operation Procedure
Experiment setup
11

GeneReach
Results
Analytical sensitivity
Analytical specificity
Comparison of diagnostic performance
12

GeneReach
Analytical Sensitivity
NCVD RAHO 6
13

GeneReach
Analytical Specificity
NCVD RAHO 6
14

GeneReach
Comparison of Diagnostic Performance
NCVD RAHO 6
15

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

GeneReach
After ASF spread in Asia…
Biosecurity
Cleaning
Disinfection
Detection
Check
the pigs
by PCR
Pulling
bad
teeth
Biosecurity Keep production
17

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

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

GeneReach
GeneReach
Thank You for the Attention

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Chandrima Spa Ajman is one of the leading Massage Center in Ajman, which is open 24 hours exclusively for men. Being one of the most affordable Spa in Ajman, we offer Body to Body massage, Kerala Massage, Malayali Massage, Indian Massage, Pakistani Massage Russian massage, Thai massage, Swedish massage, Hot Stone Massage, Deep Tissue Massage, and many more. Indulge in the ultimate massage experience and book your appointment today. We are confident that you will leave our Massage spa feeling refreshed, rejuvenated, and ready to take on the world. Visit : https://massagespaajman.com/ Call : 052 987 1315

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Chandrima Spa Ajman

We are one of the top Massage Spa Ajman Our highly skilled, experienced, and certified massage therapists from different corners of the world are committed to serving you with a soothing and relaxing experience. Luxuriate yourself at our spas in Sharjah and Ajman, which are indeed enriched with an ambiance of relaxation and tranquility. We could confidently claim that we are one of the most affordable Spa Ajman and Sharjah as well, where you can book the massage session of your choice for just 99 AED at any time as we are open 24 hours a day, 7 days a week. Visit : https://massagespaajman.com/ Call : 052 987 1315

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Chandrima Spa Ajman

The best massage spa Ajman is Chandrima Spa Ajman, which was founded in 2023 and is exclusively for men 24 hours a day. As of right now, our parent firm has been providing massage services to over 50,000+ clients in Ajman for the past 10 years. It has about 8+ branches. This demonstrates that Chandrima Spa Ajman is among the most reasonably priced spas in Ajman and the ideal place to unwind and rejuvenate. We provide a wide range of Spa massage treatments, including Indian, Pakistani, Kerala, Malayali, and body-to-body massages. Numerous massage techniques are available, including deep tissue, Swedish, Thai, Russian, and hot stone massages. Our massage therapists produce genuinely unique treatments that generate a revitalized sense of inner serenely by fusing modern techniques, the cleanest natural substances, and traditional holistic therapists.

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GEMMA Wean is a high end larval co-feeding and weaning diet aimed at Artemia optimisation and is fortified with a high level of proteins and phospholipids. GEMMA Wean provides the early weaned juveniles with dedicated fish nutrition and is an ideal follow on from GEMMA Micro or Artemia. GEMMA Wean has an optimised nutritional balance and physical quality so that it flows more freely and spreads readily on the water surface. The balance of phospholipid classes together with the production technology based on a low temperature extrusion process improve the physical aspect of the pellets while still retaining the high phospholipid content. GEMMA Wean is available in 0.1mm, 0.2mm and 0.3mm. There is also a 0.5mm micro-pellet, GEMMA Wean Diamond, which covers the early nursery stage from post-weaning to pre-growing.

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Join us for a comprehensive 90-minute lesson designed specifically for Compliance Officers and Practice/Business Managers. This 2024 HIPAA Training session will guide you through the critical steps needed to ensure your practice is fully prepared for upcoming audits. Key updates and significant changes under the Omnibus Rule will be covered, along with the latest applicable updates for 2024. Key Areas Covered: Texting and Email Communication: Understand the compliance requirements for electronic communication. Encryption Standards: Learn what is necessary and what is overhyped. Medical Messaging and Voice Data: Ensure secure handling of sensitive information. IT Risk Factors: Identify and mitigate risks related to your IT infrastructure. Why Attend: Expert Instructor: Brian Tuttle, with over 20 years in Health IT and Compliance Consulting, brings invaluable experience and knowledge, including insights from over 1000 risk assessments and direct dealings with Office of Civil Rights HIPAA auditors. Actionable Insights: Receive practical advice on preparing for audits and avoiding common mistakes. Clarity on Compliance: Clear up misconceptions and understand the reality of HIPAA regulations. Ensure your compliance strategy is up-to-date and effective. Enroll now and be prepared for the 2024 HIPAA audits. Enroll Now to secure your spot in this crucial training session and ensure your HIPAA compliance is robust and audit-ready. https://conferencepanel.com/conference/hipaa-training-for-the-compliance-officer-2024-updates

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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

13. GeneReach Analytical Sensitivity NCVD RAHO 6 13

14. GeneReach Analytical Specificity NCVD RAHO 6 14

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

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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