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virotype BVDV RT-PCR Kit

For safe detection of RNA from bovine viral diarrhea virus

  • Rapid and specific detection of RNA from bovine viral diarrhea virus
  • Amplification control included in PCR mix
  • RT-PCR run time of less than 2.5 hours
  • Can be performed in one cycler run with other virotype RT-PCR assays



The virotype BVDV RT-PCR Kit allows the safe detection of RNA from bovine viral diarrhea virus genotypes 1 and 2 (BVDV-1 and BVDV-2) in samples from cattle. The kit contains all of the reagents for real-time RT-PCR. It can be used with pools of up to 50 individual samples of blood, plasma, or serum, 25 individual samples of ear tissue, and 100 individual samples of milk.
Cat No./ID: 280373
virotype BVDV RT-PCR Kit (24)
For 24 reactions: PCR Mix, Enzyme Mix, Positive Control, Negative Control
Cat No./ID: 280375
virotype BVDV RT-PCR Kit (96)
For 96 reactions: PCR Mix, Enzyme Mix, Positive Control, Negative Control
Cat No./ID: 280377
virotype BVDV RT-PCR Kit (480)
For 480 reactions: PCR Mix, Enzyme Mix, Positive Control, Negative Control

The virotype BVDV 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 451

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



Performance
virotype BVDV RT-PCR Kit detects RNA from bovine viral diarrhea virus genotypes in samples. A validation study of 5056 samples (EDTA blood, serum, plasma, ear tissue, and milk; kindly provided by state laboratories) with known BVDV status was tested with the virotype BVDV RT-PCR Kit. Of 4570 negative samples, 4568 samples tested negative, providing a diagnostic specificity with this sample set of 99.95%. Of 486 positive samples, 484 samples tested positive, providing a diagnostic sensitivity with this sample set of 99.6% (see the table Specificity and sensitivity of virotype BVDV RT-PCR Kit). In addition, samples from the German BVD ring trial were tested with the virotype BVDV RT-PCR Kit (conducted by the Friedrich-Loeffler-Institut). The sample set contained positive, weak positive, and negative serum, as well as milk and ear tissue samples. The virotype BVDV RT-PCR kit showed excellent sensitivity detecting all positive samples including the weak positive samples (see the table BVD 2011 ring trial results).

Specificity and sensitivity of virotype BVDV RT-PCR Kit
Sample   Reference assay  virotype BVDV RT-PCR Kit result
Positive  Negative  Positive  Negative  False positive  False negative 
5056   486  4570  484  4568  2  2

Using a mixture of sample types, the virotype BVDV RT-PCR Kit has with this sample set a diagnostic specificity of 99.95% and a diagnostic sensitivity of 99.6%.

BVD 2011 ring trial results
Sample origin Expected result  virotype BDVD RT-PCR Kit CT virotype BDVD RT-PCR Kit result  
Serum/PI, BVDV-1  Positive  22.71 Positive 
Serum/TI, BVDV-1  Positive  28.80 Positive 
Serum diagnostic gap, BVDV-1  Positive  30.37 Positive 
Serum/PI, BVDV-1  Positive  23.45 Positive 
Serum/TI, BVDV-1  Positive  30.30 Positive 
Serum diagnostic gap, BVDV-1  Positive  30.94 Positive 
Serum/PI, BVDV-1  Positive  26.21 Positive 
Serum/PI, BVDV-1  Positive  24.92 Positive 
Serum/fetal calf, "HoBi"  Positive  31.43 Positive 
Milk/PI  Positive  18.71 Positive 
Milk/PI Milk 1:100 diluted  Positive  30.45 Positive 
Ear tissue/PI (Caisley tube)  Positive  27.63 Positive 
Ear tissue/PI (Caisley tube)  Positive  25.08 Positive 
Ear tissue/PI (Caisley tube)  Positive  29.28 Positive 
Ear tissue/PI (Allflex tube)  Positive  24.44 Positive 
Ear tissue/PI (Allflex tube)  Positive  25.25 Positive 
Ear tissue (Caisley tube)  Positive  23.42 Positive 

In an independent study conducted by the Friedrich-Loeffler-Institut, the virotype BVDV RT-PCR Kit showed excellent sensitivity detecting all positive samples including the weak positive samples.
Principle

This is a real-time RT-PCR kit for the detection of RNA from bovine viral diarrhea virus. 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 enzyme mix 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, plasma, serum, ear tissue, or milk) using a suitable sample preparation kit. The virotype Tissue Lysis Reagent provides a fast, easy-to-use, and convenient method for RNA preparation from ear tissue samples. The extracted RNA is used for the RT-PCR, which is run on a thermal cycler.

Applications

The virotype BVDV RT-PCR Kit is for detection of RNA from bovine viral diarrhea virus (BVDV) in samples from cattle.

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