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virotype Influenza A RT-PCR Kit

For detection of RNA from influenza A virus in samples from birds and swine

  • Rapid and safe detection of RNA from all 16 HA and 9 NA subtypes of influenza A virus, including H5N8
  • Easy-to-use single reaction mix containing internal control
  • RT-PCR run time of less than 2 hours
  • Can be performed in one cycler run with other virotype RT-PCR assays


The virotype Influenza A RT-PCR Kit allows the safe detection of RNA from influenza A virus in samples from birds and pigs. The kit contains all of the reagents for real-time RT-PCR in a single ready-to-use reaction mix. It can be used with oropharyngeal, tracheal, and cloacal swabs, fecal samples, or tissue samples from birds, and nasal swabs, bronchoalveolar lavage fluid (BALF), and tissue samples from swine. It can also be used with cell culture supernatants. Due to the high sensitivity of the test, pools of up to 10 individual swab samples can be tested.
Cat No./ID: 282603
virotype Influenza A RT-PCR Kit (24)
For 24 reactions: Master Mix, Positive Control, Negative Control
Cat No./ID: 282605
virotype Influenza A RT-PCR Kit (96)
For 96 reactions: Master Mix, Positive Control, Negative Control

The virotype Influenza A 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 538



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Sensitive detection of influenza A RNA.

Using the virotype Influenza A RT-PCR Kit, real-time RT-PCR was performed in triplicate on a titration series of in vitro RNA (2 x 107 copies/μl). There is a linear correlation between the log of the RNA copy number and the amplification results (CT). The correlation coefficient is 1.0.

Performance
virotype Influenza A RT-PCR Kit detects RNA from Influenza A virus. A validation study of 343 samples, with known influenza A status (186 positive and 157 negative; kindly provided by the OIE and National Reference Laboratory for Avian Influenza / Fowl Plague [AI] and private laboratories), was tested with the virotype Influenza A RT-PCR Kit. Of 186 positive samples, 178 samples tested positive, providing a diagnostic sensitivity of 96%. In this study all negative samples tested negative, providing a diagnostic specificity of 100%. In addition, a titration series was performed with ten-fold dilutions from 108 to 10 copies/well of influenza A RNA. The results demonstrate a high correlation (99%) between the number of copies and the amount of amplification product in the range from 108 to 100 copies/well, and positive detection of the virus down to 10 copies/well (see the figure Sensitive detection of Influenza A RNA).
Principle

This is a real-time RT-PCR kit for the detection of RNA from influenza A 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.

The virotype Influenza A RT-PCR Kit uses two specific primer/probe combinations, one for influenza A virus RNA and one for the control RNA.

Procedure

The kit contains a ready-to-use mix with all of the necessary reagents for the detection of influenza A virus 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 starting material 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 Influenza A RT-PCR Kit is for detection of RNA from influenza A virus in in samples from birds and swine.

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