Validation of African swine fever real-time PCR Kit

A new duplex real-time PCR kit that targets the p72 gene and an internal control has been developed and assessed for the diagnosis of ASFV.
calendar icon 25 October 2016
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Thermo Fisher ASFV Diagnostic Kit

The disease is widely endemic in many parts of Africa, of Southern Europe and increasingly becoming a threat in Eastern Europe. As there is still no vaccine or treatment available, monitoring and controlling of the disease by means of diagnosis is the only way to control the disease and is of utmost importance. A duplex real-time PCR kit targeting the p72 gene and an internal control has been developed and its performance for diagnosis of ASFV has been assessed.

In order to demonstrate the sensitivity and specificity of the new LSI VetMAX™ African Swine Fever Virus detection kit, different internal and field studies including animal infection experiments were carried out (INIA, Spain; CVI, Netherlands; Germany). 1600 negative samples from ASFV free regions (Germany & Spain) and 33 different pathogens were tested to demonstrate specificity of the assay. About 100 ASFV positive samples from Africa and Europe were also tested.

Results of the ASFV kit showed 100% sensitivity in all tested sample materials (blood, serum and tissues) and 100% specificity. No cross reaction was found with other pathogens and a serial dilution of the ASFV target sequence led to a limit of detection (LOD) of 16 genome copies per PCR reaction. The experimental LOD was 5x103 copies per mL in serum and 1x104 copies per mL in blood.

The LSI VetMAX™ African Swine Fever Virus detection kit fulfills all the validation criteria of PCR characteristics and complete method, as required by the NF U47-600-2 standard.

Early Detection of ASF Virus

African Swine Fever (ASF) is a DNA virus from the Asfaviridae Family. ASFV infects all Suidae (domestic and wild animals) but is not a human health threat. The virus is found in all body fluids and tissues of infected pigs. They usually become infected by direct contact with sick animals or by ingestion of infected products.

ASFV is highly resistant in the environment. ASF disease is characterized by high fever, loss of appetite, haemorrhages in the skin and internal organs and death can occur within 2 to 10 days on average. ASF cannot be differentiated from classical swine fever by either clinical or post-mortem examination. It is an economically important disease that is widely endemic in many parts of Africa and that has become a real threat in Eastern Europe. In order to improve ASF diagnosis, a new duplex real time PCR kit was developed.


ASFV Distribution January - June 2016

ASFV Distribution July - October 2016

Further Reading

You can view the full research poster by clicking here.

For more information about swine diagnostics, click here or connect to the Thermo Fisher Scientific Swine Resource Center.

Sarah Mikesell


Sarah Mikesell grew up on a five-generation family farming operation in Ohio, USA, where her family still farms. She feels extraordinarily lucky to get to do what she loves - write about livestock and crop agriculture. You can find her on Twitter or LinkedIn.

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