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virotype BTV RT-PCR Kit

For detection of RNA from all bluetongue virus serotypes plus Toggenburg virus in samples from ruminants

  • Rapid and reliable detection of RNA from bluetongue virus
  • Detection of all 25 known BTV serotypes, including Toggenburg virus
  • Amplification control included in PCR mix
  • RT-PCR run time of less than 2 hours
  • Can be performed in one cycler run with other virotype RT-PCR assays


The virotype BTV RT-PCR Kit allows the safe detection of RNA from bluetongue virus (BTV) in samples from cattle, sheep, and goats. The kit contains all of the reagents for real-time RT-PCR. It can be used with pools of up to 10 individual samples of blood and with tissue samples from the spleen and lymph nodes.
Cat No./ID: 280433
virotype BTV RT-PCR Kit (24)
For 24 reactions: PCR Mix, Enzyme Mix, Positive Control, Negative Control
Cat No./ID: 280435
virotype BTV RT-PCR Kit (96)
For 96 reactions: PCR Mix, Enzyme Mix, Positive Control, Negative Control

The virotype BTV RT-PCR Kit is for veterinary use only.

In Germany: Registered in accordance with §17c of the German Law on Animal Diseases. Registration No.: FLI-B 482

In Switzerland: Approved by the Federal Veterinary Office (BVET)



Performance
virotype BTV RT-PCR Kit detects RNA from bluetongue virus in samples. A validation study of 400 bovine and ovine blood samples, with known BTV status (233 positive and 167 negative; kindly provided by the state laboratories), was carried out with the virotype BTV RT-PCR Kit. Of 167 negative samples, 1 sample tested positive, providing a diagnostic specificity of 99.4%. Of 233 positive samples, 1 sample tested negative, giving a diagnostic sensitivity of 99.6%. The kit can detect as few as 10 copies of BTV RNA per sample. A titration series was performed with ten-fold dilutions from 107 to 10 copies/well of BTV RNA. The results demonstrate a high correlation (99%) between the number of copies and the amount of amplification product in the range from 107 to 100 copies/well, and positive detection of the virus down to 10 copies/well (see the figure Sensitive detection of BTV RNA).
Principle

This is a real-time RT-PCR kit for the detection of RNA from bluetongue virus, the causative agent of the highly infectious bluetongue disease, which mainly affects cattle, sheep, and goats.

RT-PCR is based on the amplification of specific regions of the pathogen genome. It is a sensitive and specific tool for pathogen detection.

Real-time RT-PCR involves the reverse transcription of viral RNA and subsequent amplification of the cDNA by PCR. The amplified product is detected using fluorescent dyes. These are usually linked to oligonucleotide probes that bind specifically to the amplified product. Monitoring the fluorescence intensities during the PCR run (i.e., in real time) allows the detection of the accumulating product without the need to re-open the reaction tubes afterward.

Procedure

The kit contains a ready-to-use PCR mix and an enzyme mix with all of the necessary reagents for the detection of BTV RNA and the required positive and negative controls. With this kit, both reverse transcription and PCR are performed in one reaction tube, reducing the risk of contamination.

RNA is extracted from the sample material (blood or tissue) using a suitable sample preparation kit. The extracted RNA is used for the RT-PCR, which is run on a thermal cycler.

Applications

The virotype BTV RT-PCR Kit is for detection of RNA from bluetongue virus (BTV) in samples of from cattle, sheep, and goats.

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