African swine fever (ASF) is an acute, severe, and highly contagious infectious disease caused by the African swine fever virus (ASFV) infecting domestic pigs or wild boars. Its morbidity and mortality can reach 100%, seriously endangering the global pig industry and causing incalculable economic losses. The World Organization for Animal Health (OIE) lists it as a reportable animal disease.

At present, there is no vaccine or effective treatment for African swine fever. Rapid and accurate diagnosis of African swine fever through laboratory diagnostic methods is crucial to preventing the spread of the disease. qPCR as the main tool for routine diagnosis of ASF is recommended by OIE.

Yeasen is a manufacturer with rich experience in IVD raw materials R&D. In the face of African swine fever, yeasen responds positively and cooperates with customers to develop three whole process solutions from extraction to detection of African swine fever virus. So what are three solutions? And what are applications of these solutions? Continuing to read...

Figure 1. Yeasen African Swine Fever detection solutions

 

Solution I: TaqMan Multiplex qPCR Master Mix (UDG plus)（inquiry）

Product Features

👍UDG anti-pollution system, effectively preventing the risk of aerosol contamination

👍Long-term stability of pre-mixed PCR solution(master mix + primers + probes) at room-temperature, greatly simplifying the detection difficulty

👍High amplification efficiency and good repeatability

👍Suitable for multi-platform and multi-system

Case Show

👉Long-term stability of pre-mixed PCR solution
Cat#13171 was used to configure the pre-mixed PCR solution that can be placed at 37°C for 7 days, and repeated freezing and thawing 15 times in the amplification of the ASF plasmid system. The results showed the Ct value and sensitivity of PCR solution has not significantly differ after placing 7 days at 37℃, and the Ct value did not change significantly after repeated freezing and thawing 15 times.

Figure 2. Cat#13171 TaqMan Multiplex qPCR Master Mix (UDG plus)(Inquire) used in pre-mixed PCR solution with long-term stability in amplification of ASF plasmid system. 2A-Ct value at 37℃ for 0 day and 7 days 2B-Sensitivity at 37℃ for 0 day and 7 days 2C-Stability after freeze-thaw 15 times

👉Stable and Reproducible reagents
Four different batches of Cat#13171 were selected to repeat the ASFV plasmid system amplification experiment 20 times. There were no significant differences in Ct values, with a CV of less than 1%.

Figure 3. Repeatability test of Yeasen Cat#13171

👉Multi-platform multi-system applicable
Yeasen Cat#13171 was used to amplify African swine fever ASFV/ACT dual plasmids on Bio-Rad CFX96, ABI Q5 and Slan platforms respectively. It can be seen that Yeasen Cat#13171 is suitable for various types of instruments, and the data is reliable and stable.

Figure 4. Bio-Rad CFX96 Real-Time PCR System

Figure 5. ABI Q5 Real-Time PCR System

Figure 6. Slan Real-Time PCR System

 

Solution II:Universal TaqMan multiplex qPCR master mix

Yeasen's Universal TaqMan multiplex qPCR master mix (Cat#11211) is deeply optimized for efficient amplification of low-concentration templates using antibody-modified Hot-Start Taq DNA polymerase and matched multi-plex buffer. This product supports up to 4-plex PCR reactions in single tube and is ideal for genotyping and multiplex qPCR analysis.

Product Features

👍Strong tolerance to blood impurities, can be used with Yeasen nucleic acid extraction reagent Cat#13720 Nucleic Acid Release Reagent V2, ideal for direct qPCR
👍Ultra-high sensitivity to 3 copies/reaction, greatly improve detection accuracy
👍Long-term stability of pre-mixed PCR solution (master mix + primers + probes) at room-temperature, greatly simplifying the detection difficulty

Case Show

👉Strong tolerance to blood impurities
Compared with similar products from other brands, Yeasen Cat#11211 has a higher tolerance to inhibitors in blood samples. The results showed that Yeasen Cat#11211 has a high tolerance to EDTA and sodium citrate, and can withstand 0.5% whole blood.

Figure 7. Cat#11211 Universal TaqMan multiplex qPCR master mix and master mix from various brands are used for ASF qPCR solutions supplemented with 3 mM EDTA (3A), 6 mM sodium citrate (3B), 0.5% blood (3C), respectively.

👉Ultra-high sensitivity to 3 copies/reaction

The amplification of the ASFV plasmid with 20 replicates was performed based on the products from different brands. The results of experiments show that the detection rate of Yeasen products is higher.

Figure 8. Comparison of detection rates among similar products from different brands (3 copies in final system)

👉Long-term stability, stable for 7 days at room temperature

The PCR solution of ASFV/ACT dual plasmid system can maintain stable performance after 7 days at 4℃, 25℃ and 37℃. It can be seen that the Ct value and the fluorescence value did not change significantly.

Figure 9. Stability of PCR solution

Solution III: Isothermal Amplification

Yeasen's products support the isothermal amplification technology that helps perform ASF detection faster and more efficiently. Cat# 14402 Hieff™ Bst Plus DNA Polymerase (40 U/μL) is derived from Thermophilic Geobacillus sp DNA Polymerase I, lacking 5'-3' exonuclease activity. This product has stronger 5'-3' DNA polymerase activity, strand displacement activity and dUTP tolerance, which is more suitable for anti-pollution isothermal amplification reactions, such as LAMP, CPA, etc.

Product Features

👍High amplification efficiency

👍High Yield Amplification

👍dUTP tolerance, can be used in combination with Cat#10303 Uracil DNA Glycosylase (UDG/UNG)(Inquire), heat-labile, 1 U/μL to prevent the risk of aerosol contamination

Case Show

👉Superior alternative to imported brand N* products 

The amplification process was performed using Bst DNA polymerase from Yeasen and N* Company, respectively. There was no significant difference in Ct value and fluorescence intensity between different brands.

Figure 10. Amplification curves of different brands

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