Duplex ASFV Fast qPCR Kit

cat. no. E0473

Certified for veterinary use

Description

  • Duplex ASFV Fast qPCR Kit is intended for the detection of DNA from African swine fever virus (ASFV) in blood, serum, plasma, swab samples (from mouth, nose, nasopharynx), tissues and organs of pigs and wild boars.
  • The high sensitivity of the kit allows the detection of the ASF virus in both single samples and mixed (pooled) samples consisting of a maximum of 5 individual samples.
  • Duplex ASFV Fast qPCR Kit allows to use a standard or fast PCR amplification program, depending on the user’s preferences and the type of real-time PCR device used.
  • The kit contains specific primers for the highly conserved ASFV p72 gene fragment. The amplified p72 gene fragment is detected using a probe labeled with FAM dye.
  • Additionally, the kit contains an endogenous internal control consisting of primers and a probe labeled with HEX dye for a fragment of the porcine ACTB gene encoding beta-actin. Internal control allows both extraction and amplification to be monitored.
  • DNA used for PCR-based diagnostics should be of good quality and purified using dedicated, commercial kits using silica or magnetic bead technology.
  • Duplex ASFV Fast Master Mix contains all of the necessary reagents, including  Perpetual Taq DNA polymerase, reaction buffer, deoxynucletide triphosphates (dNTPs), primers and probes combination and passive reference dye (ROX).
  • Perpetual Taq DNA Polymerase is a modified “hot start” enzyme that is blocked at moderate temperatures by antibodies and allows room temperature reaction setup. The polymerase activity is restored during the initial 2-minutes denaturation step.
  • The passive ROX reference dye included in the master mix enables fluorescence normalization in particular cyclers. The use of ROX dye is required for all Applied Biosystems real-time PCR cyclers and optional for Agilent cyclers. ROX compensates for differences in fluorescence signal between wells caused by small differences in reaction volume and fluorescence fluctuations. ROX is not involved in the PCR reaction and does not interfere with real-time PCR on any instrument.
  • Positive control – PC contains both DNA fragment ASF virus and the porcine ACTB gene and serves to prove of the kit’s operation, including the correct preparation of the PCR reaction. The fluorescence signal for ASF virus DNA in the FAM channel has CT= 29±2, and for the ACTB gene in the HEX channel has CT= 29±2.
  • A complementary product to the Duplex ASFV Fast qPCR Kit (which can be ordered additionally) is the ASFV Positive Extraction Control (cat. no. E0474). The ASFV Positive Extraction Control is a reference solution containing a synthetic DNA fragment of the ASF virus and a synthetic fragment of the porcine ACTB gene and can be used as a positive control for the isolation of the ASF virus.