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virotype ASFV PCR Kit

For detection of DNA from African swine fever virus in swine samples

  • Duplex real-time PCR for rapid and specific detection of ASFV DNA
  • Easy-to-use reaction mix containing controls
  • PCR run time of less than 2 hours
  • Can be performed in one cycler run with virotype CSFV RT-PCR Kit

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.
Cat No./ID: 281805
virotype CSFV RT-PCR Kit (96)
For 96 reactions: CSFV Mix, Positive Control, Negative Control
Cat No./ID: 281903
virotype ASFV PCR Kit  (24)
For 24 reactions: Master Mix, Positive Control, Negative Control
Cat No./ID: 281905
virotype ASFV PCR Kit  (96)
For 96 reactions: Master Mix, Positive Control, Negative Control
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).
Performance
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.


Table 1. Sensitive detection using the virotype ASFV PCR Kit
AFSV strain   Species                   Days post infection Sample type                   FLI virotype
ASFV protocol
FAM
 virotype
ASFV protocol
HEX
virotype
ASFV/CSFV protocol
FAM
 virotype
ASFV/CSFV protocol
HEX
 Sardinia      Wild boar-1  8 Blood 21.10 18.99  22.59 19.86  22.45
 Sardinia  Wild boar-1  8 Bone marrow 21.73 19.59  22.34 19.17  22.64
 Sardinia  Wild boar-1  8 Fecal 29.34 27.42  24.27 27.56  24.44
 Sardinia  Wild boar-1  8 Lung 24.01 20.62  22.20 20.42  22.50
 Sardinia  Wild boar-1  8 Lymph node 24.47 21.90  21.40 23.22  21.38
 Sardinia  Wild boar-1  8 Oral-pharyngeal 33.40 31.09  29.10 31.20  29.37
 Sardinia  Wild boar-1  8 Salivary gland 25.69 23.78  24.33 23.60  24.68
 Sardinia  Wild boar-1  8 Tonsil 23.46 21.24  21.14 21.11  21.25
 Armenia  Wild boar-2  8 Blood 16.22 16.84  20.91 18.54  20.90
 Armenia  Wild boar-2  8 Oral-pharyngeal no CT 34.92  28.39 36.42  28.18
 Kenia05  Pig-1  14 Blood 21.16 19.47  23.07 19.32  23.65
 Kenia05  Pig-1  14 Bone marrow 20.04 16.42  22.09 16.14  22.59
 Kenia05  Pig-1  14 Fecal 26.66 25.67  24.98 25.73  25.37
 Kenia05  Pig-1  14 Lung 25.48 21.83  23.36 21.53  23.69
 Kenia05  Pig-1  14 Lymph node 21.22 18.10  20.25 17.81  20.43



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