High Pure Viral Nucleic Acid Kit
Low to medium throughput viral nucleic acid isolation.
For general laboratory use.
|Product No.||Pack Size|
To order these products, please contact your Roche order management team.
The High Pure Viral Nucleic Acid Kit is intended for general laboratory use and efficiently purifies viral nucleic acids from serum, plasma, or whole blood using sample volumes of up to 200 μL. When using whole blood as starting material, total nucleic acids are isolated, including viral nucleic acids. The purified viral nucleic acid is eluted in nuclease-free water, and is suitable for direct use in PCR and RT-PCR.
- Isolate both RNA and DNA, allowing simultaneous detection of both types of virus.
- Remove inhibitors that might interfere with downstream assays, ensuring greater assay specificity, sensitivity, and reproducibility (see Figure 1).
- Obtain a concentrated sample, suitable for direct assay (no precipitation required).
- Scale the procedure to accommodate samples with low viral loads.
- Save time with a kit that prepares multiple PCR/RT-PCR templates in just minutes.
- Eliminate the use of hazardous organic compounds such as cesium chloride, phenol, chloroform, and ethidium bromide.
- Binding Buffer
- Poly (A), lyophilized
- Proteinase K, recombinant, lyophilizate
- Inhibitor Removal Buffer
- Wash Buffer
- Elution Buffer
- High Pure Spin Filter Tubes (containing glass fiber fleece)
- Collection Tubes
The High Pure Viral Nucleic Acid Kit isolates highly purified viral nucleic acids (DNA and RNA) from mammalian plasma, serum, or whole blood. The purified viral nucleic acid may be used directly as templates for standard or long-template PCR.
Figure 1: Sensitivity of viral RNA detection.
For research purposes, serial dilutions of serum samples containing HGV, HCV, and HIV were processed according to the kit protocol. Viral nucleic acid from each sample was eluted in a total volume of 100 µL. Aliquots (20 µL each) from the eluates were analyzed by one-step RT-PCR, using either Tth DNA polymerase (for HCV and HIV) or a mixture of AMV reverse transcriptase and Taq DNA polymerase (for HGV). PCR products were quantified by electrochemiluminescence, using a biotinylated capture probe and a ruthenium-labeled detection probe.
Result: Assuming the kit recovers all the virus from each original sample, the assays could detect as few as 20 – 40 viral copies in each of the isolated samples.
Nucleic acids bind to the surface of the glass fiber fleece in the presence of a chaotropic salt (guanidine HCl). This allows the High Pure Filter Tube to specifically immobilize nucleic acids (both DNA and RNA) while they are freed of contaminants.
Note: A special Inhibitor Removal Buffer is included which allows the use of heparinized sample material. This buffer increases the sensitivity and reproducibility of assays performed with the isolated viral nucleic acid, even when the sample contains heparin.
Capacity: The High Pure Spin Filter Tubes hold up to 700 μL sample volume.
Typical Nucleic Acid Recovery
Starting Material and Quantity
Number of Reactions
|Serum, plasma, whole blood, supernatant from
cell culture, 200 - 600 µL
|Product detectable by PCR or RT-PCR||20 minutes||100/200 µL|
As a prerequisite for PCR or RT-PCR, viral nucleic acids must be isolated from serum, plasma, or whole blood. Viral lysis is accomplished by incubating the sample in a special Lysis/Binding Buffer in the presence of Proteinase K. Sample is mixed with a chaotropic salt and applied to the glass fiber fleece in a High Pure Spin Filter Tube. Under the buffer conditions used in the procedure, the viral nucleic acids bind to the glass fleece, while contaminating substances (salts, proteins, nucleotides, mineral oil, and other contaminants) do not. Brief wash-and-spin steps readily remove these contaminants. Once purified, the viral nucleic acid can be easily eluted in a small volume of low-salt buffer or water.