African Swine Fever (ASFV) Virus Real-Time PCR Detection Kit

Target and Hazard: African swine fever (ASF) is caused by African swine fever virus (ASFV, family Asfarviridae). ASFV is a double-stranded DNA virus with exceptional resistance in the environment, surviving for long periods in blood, tissues, meat products, manure and contaminated materials, so slaughter, processing, transport and feed waste can all form links in the transmission chain. Pigs are the only susceptible animals; infected pigs develop high fever, cyanosis, vomiting and bloody diarrhoea, mortality in the acute form approaches 100%, and no safe and effective commercial vaccine is available anywhere in the world. The disease is a Class I animal disease in China and a WOAH-listed disease requiring notification: once confirmed it requires quarantine, culling and safe disposal.

Significance of Testing: This kit detects ASFV nucleic acid directly and is a core laboratory method for early confirmation of African swine fever: it can detect the virus very early in infection, before seroconversion and even before typical clinical signs appear, and the Ct value gives a rough indication of viral load. It is used for outbreak confirmation and review of suspected cases, investigation of virus-carrying pigs and contaminated environments, and laboratory confirmation in movement, slaughter and carcass-disposal settings. Combined with colloidal gold antigen cards it forms a complete 'on-site screening – laboratory confirmation' chain. Note that, because no vaccine is available, ASFV antibody testing serves only epidemiological investigation and infection tracing and cannot be used to assess immune protection; this kit does not serve that purpose.

Principle: Specific primers and a TaqMan probe are designed against highly conserved regions of the ASFV genome; probe hydrolysis during amplification generates fluorescence that accumulates with the product, and results are interpreted by real-time monitoring of the amplification curve together with the Ct value. A UNG-dUTP anti-contamination system is included to reduce the risk of aerosol carry-over.

Performance and Applications: Supplied with nucleic acid extraction reagents; whole blood, tissue and environmental samples can be tested in batches within about 2–3 hours; a real-time PCR instrument is required and results are read from the amplification curve and Ct value; store frozen at -20 °C or below, protected from light, shelf life 12 months. Intended for laboratory confirmation and pathogen surveillance on large-scale pig farms, at animal disease prevention and control institutions and third-party testing laboratories, and in slaughter and animal movement settings. A positive result must be reported immediately in accordance with national regulations and the samples handled appropriately. Method: TaqMan probe real-time fluorescent quantitative PCR | Samples: whole blood, serum, plasma, spleen/lymph node/tonsil homogenate, environmental swabs | Time to result: ~2–3 h including nucleic acid extraction | Reading: real-time PCR instrument, amplification curve and Ct value | Anti-contamination design: UNG-dUTP system included | Format: as stated on the label | Storage: -18 °C, protected from light; avoid repeated freeze-thaw cycles; shelf life 12 months | Note: limit of detection, specificity and agreement with virus isolation/sequencing are subject to our internal QC report.