African swine fever (ASF) is a highly contagious and often fatal viral disease affecting domestic and wild pigs. It is caused by African swine fever virus (ASFV), a large double-stranded DNA virus.
ASF was first reported in Kenya in 1921 and is historically associated with a sylvatic cycle involving wild suids and soft ticks. Today, ASF remains a major global animal health concern. The World Organisation for Animal Health (WOAH) classifies ASF as a notifiable animal disease, while it is classified as a Category I animal disease in China.
ASFV can have an incubation period of approximately 4–19 days, and highly virulent strains can cause mortality approaching 100%. Currently, there is no widely available, safe, and effective commercial vaccine or specific antiviral treatment.
Therefore, early and reliable ASFV detection is critical for disease surveillance, outbreak control, and biosecurity management.
Key Challenges in Animal Disease Molecular Detection
Animal disease molecular testing must address several practical challenges:
- Complex Sample Matrices: Blood, feces, and tissue may contain PCR inhibitors that affect amplification.
- Low-Abundance Targets: Early-stage infections may contain limited pathogen nucleic acids, requiring sensitive detection.
- High Sample Throughput: Surveillance programs may involve hundreds or thousands of samples, requiring scalable workflows.
- Contamination Control: High-volume PCR testing increases the risk of carryover contamination.
- Reagent Stability: Decentralized testing requires reagents that are easier to transport and store.
These challenges make sample preparation, amplification chemistry, workflow design, and reagent stability equally important for reliable testing.
Figure 1. Structural morphology of African Swine Fever Virus (ASFV).
Common Methods for ASFV Detection
Several molecular technologies can be used for African swine fever virus detection. The appropriate method depends on the testing scenario, required sensitivity, instrumentation, and workflow. Among these methods, real-time PCR is a key technology for ASFV molecular detection because of its sensitivity, specificity, and compatibility with standardized laboratory workflows.
The B646L gene, encoding the major capsid protein p72, is one of the most commonly used ASFV detection targets. Other targets, such as CD2v and MGF 360–505R, may also be used depending on the assay purpose.
Importantly, detection performance depends not only on the target gene but also on the primer/probe design, DNA polymerase, reaction chemistry, and sample matrix.
Yeasen ASFV Detection Solutions
To support different ASFV molecular testing workflows, Yeasen provides solutions covering qPCR, LAMP, RPA, and customized diagnostic formulations.
qPCR Solution
Different ASFV assays may use different primer/probe systems. To improve assay compatibility, Yeasen has screened multiple Taq DNA polymerases and optimized different qPCR formulations for molecular detection applications.
- Universal qPCR Solution —Hieff Unicon™ HiSens TaqMan Multiplex qPCR Mix (UDG plus) (Cat#16967)
A versatile qPCR master mix for routine ASFV molecular detection and assay development, offering broad compatibility with different primer/probe systems.
Figure 2. Performance comparison of three qPCR master mixes on 45 ASFV clinical samples.
The 16967 master mix achieves the highest positive detection rate (91.11%) and the lowest mean Ct value, demonstrating superior sensitivity for clinical specimens.
Figure 3. Ct value distribution of ASFV positive clinical samples detected with three qPCR master mixes.
Each dot represents one positive sample, and horizontal lines denote the mean Ct of each group. The 16967 reagent detects more weak-positive samples at high Ct range (>35), showing superior performance on low viral load specimens.
- Direct Swab qPCR Solution— Hieff Unicon™ Robust Multiplex qPCR PreMix V1 (UDG Plus) (Cat#16945)
Designed for direct detection from swab samples, helping simplify sample preparation and streamline ASFV screening workflows.
Figure 4. Direct amplification from swab samples: 16945 vs. comparable market products.
Pharyngeal swabs were processed in Nucleic Acid Release Reagent (Cat#16964) or TE buffer to dilute plasmid templates. 16945 demonstrated superior inhibitor tolerance compared to market alternatives, confirming its suitability for direct swab amplification.
Customer Feedback:
Figure 5. Stability and clinical performance of Yeasen 16945 vs Supplier V*.
Case1: Accelerated stability test of ASFV & LSDV assays. Case2: Real ASFV sample testing. Yeasen 16945 showed lower average Ct and recovered a weak-positive sample undetected by Supplier V*, demonstrating superior stability and sensitivity.
Isothermal Amplification Solution
For rapid and simplified ASFV molecular testing, Yeasen offers a robust LAMP-based isothermal amplification solution. Designed for high efficiency and ease of use, our LAMP reagents empower rapid screening, decentralized testing, and field-oriented molecular detection.
- RT-LAMP Fluorescent Assay Kit(Bst Plus)(Cat#16729)
Figure 6. Compatibility and Performance Evaluation of Dye-Based LAMP Reagent with Nucleic Acid Release Reagent
(A) Direct LAMP amplification from oral swab lysates. Oral swabs were processed in Nucleic Acid Release Reagent (Cat#16964) and used directly as templates. Results demonstrate effective amplification when paired with the dye-based LAMP Reagent (Cat#16729ES).
(B) ASFV target detection: Nucleic Acid Release Reagent vs. standard DNA diluent. ASFV targets diluted in Nucleic Acid Release Reagent (Cat#16964) and standard DNA diluent were tested at identical concentrations. Results indicate that the Nucleic Acid Release Reagent does not compromise target detection.
Customized Solutions
Yeasen provides customized solutions for liquid and lyophilized molecular diagnostic reagents, covering PCR, LAMP, RPA, and lyophilization technologies.
- New Product Development: Develop and optimize assays based on your targets, workflow, and performance requirements, with support for scale-up and regulatory submissions.
- Custom Enzymes: Develop or substitute key enzymes to improve performance, compatibility, or cost-efficiency.
- Custom Systems & Formulations: Customize master mixes, component concentrations, dyes, primer/probe systems, lyophilization formulations, and isothermal amplification systems.
- Custom Finished Products: Manufacture customized molecular diagnostic reagents and kits based on your formulation, process, and specifications.
From enzyme selection and assay optimization to formulation and finished products, Yeasen supports your molecular diagnostic development from R&D to commercialization.
Early detection is a critical component of African swine fever prevention and control. qPCR, LAMP, and RPA provide complementary approaches for different ASFV testing scenarios.
With a portfolio of qPCR master mixes, isothermal amplification reagents, molecular diagnostic enzymes, and customized formulation services, Yeasen supports the development of reliable ASFV molecular testing solutions—from assay development to customized diagnostic products.
Related Products
|
Product Category |
Product Function |
Product Name |
Catalog No. |
|
Sample Preparation |
Nucleic Acid Release Agent |
Nucleic Acid Release Agent V3 |
16964ES |
|
Magnetic Universal Viral DNA/RNA Extraction Kit |
18521ES |
||
|
ASFV Kits |
ASFV Detection Kit |
African Swine Fever Virus Fluorescent PCR Detection Kit (UDG plus) |
16961ES |
|
qPCR Premix (Primers & Probes Not Included) |
Rapid, Fully Formulated, ASFV Detection V2 |
16967ES |
|
|
Direct Swab Amplification, Fully Formulated, Low Carryover |
16945ES |
||
|
High Sensitivity, Universal, Lyophilizable |
Hieff Unicon™ Universal TaqMan Multiplex qPCR Master Mix (UDG plus) |
11893ES |
|
|
PCR Instrument |
Real-Time Fluorescence Quantitative PCR Instrument |
Celemetor™ Real-Time Fluorescence Quantitative PCR Analysis System |
80521ES |
|
Nucleic Acid Decontamination Agent |
Aerosol Decontamination |
DNA/RNA Remove Aerosol Decontamination Agent |
19702ES |
|
LAMP Reagent |
Fluorescent Dye Method |
16729ES |
|
|
RPA Reagent |
Fluorescent RPA (Exo Probe, Lyophilized) |
DNA Lyo Rapid Isothermal Amplification Kit (Fluorescent) |
16904ES |
|
Chromatographic RPA (Nfo Probe, Lyophilized) |
DNA Lyo Rapid Isothermal Amplification Kit (Chromatographic) |
16906ES |
|
|
Electrophoretic RPA (Lyophilized) |
DNA Lyo Rapid Isothermal Amplification Kit (Basic) |
16908ES |