virotype ASFV PCR Kit
For detection of DNA from African swine fever virus in swine samples
The virotype ASFV PCR Kit allows the safe detection of DNA from African swine fever virus (ASFV) in samples from pigs and wild boar within two hours. The kit contains all of the reagents for real-time PCR in a single ready-to-use reaction mix. It can be used with serum, plasma, EDTA-blood, tissue and swab samples from pigs and wild boar.
As recommended by the World Organisation for Animal Health (OIE) and the Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health (FLI), Germany, DNA and RNA from ASFV and CSFV respectively, can be extracted simultaneously using the QIAamp Viral RNA Mini Kit. For automated sample preparation, the QIAcube can be used. Extracted nucleic acids can then be tested in the same cycler run for both ASFV and CSFV using the virotype ASFV PCR Kit and virotype CSFV RT-PCR Kit, respectively.
The virotype ASFV PCR Kit is for veterinary use only. In Germany: Licensed in accordance with §11 of the German Animal Health Act (FLI-B 670).
QIAGEN’s virotype ASFV PCR Kit is a highly sensitive and specific solution for reliable and rapid detection of DNA from ASFV in serum, plasma, EDTA-blood, tissue and swab samples from pigs and wild boar.
In combination with the QIAamp Viral RNA Mini Kit and the virotype CSFV RT-PCR Kit, QIAGEN provides a complete and time-saving workflow solution for ASFV and CSFV detection. The performance using the virotype ASFV PCR Kit protocol or the PCR protocol for simultaneous amplification of ASFV DNA and CSFV RNA does not affect the quality of detection. The high sensitivity of the virotype ASFV PCR Kit allows early detection of the pathogen in individual, as well as in pooled samples.
Analytical sensitivity was determined by a titration series of in-vitro ASFV DNA.
The detection limit of the kit was shown to be 10 copies of ASFV DNA.
Further validation was conducted with ASFV positive pig and wild boar samples, experimentally infected with strains Armenia, Sardinia and Kenia05 (see Table 1). DNA was extracted from blood, spleen, lymph nodes, tonsils, salivary gland, oral-pharyngeal swaps and bone marrow.
When testing positive samples, the assay demonstrated high sensitivity. By testing 165 ASFV-negative samples, a specificity of 100% was determined. In addition, no cross-reactivity was detected with other porcine viral pathogens such as CSFV, porcine reproductive and respiratory syndrome virus (PRRSV), swine influenza virus (SIV) or porcine circovirus 2 (PCV2). Testing the intra- and inter-assay variance showed excellent reproducibility.
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