Instant procedure delivering RNA in minutes
ViRNAExTM provides fast purification of high-quality RNA from cells collected by standard nasopharyngeal swabbing. Samples can be conveniently delivered in classical viral transport media. A nucleic acid extract that is suitable for this PCR should be used with ViRNAExTM. Acceptable specimen types include nucleic acid extracts of nasal swabs, throat swabs, nasopharyngeal swabs, nasopharyngeal aspirate, tracheal aspirate, bronchial washing or bronchoalveolar washing.
RNA purification using the ViRNAExTM can be automated on any pipetting automat. For smaller and larger samples, the volume of extracting buffer may be adjusted. The recommended starting volume is 100 µl of sampling solution.
Introduction
ViRNAexTM RNA extraction solution enables instant and inexpensive RNA isolation from viruses with a possibility to use RNA subsequently in real time- PCR testing (Volume recommended: 5-10 µl of solution per qPCR reaction). The duration of RNA sample preparation is shortened on approx. 15 minutes including pipetting steps.
Recommended RT-PCR kit
Using CoVirED RT-PCR test for COVID-19 detection in combination with ViRNAexTM is strongly recommended. CoVirED PCR protocol includes internal PCR reaction control as well as control of human cells presence using human endogenous control gene probe. CoVirED (Corona Virus Extraction and Detection kit) is delivered for a favorable price if combined with ViRNAexTM (one package). More info here.
Package:
ViRNAExTM – 10 mL (for 100 rxns)
ViRNAExTM – 50 mL (for 500 rxns)
| Storage after opening: | under 8 °C |
| Unit Definition: | 10mL or 50mL |
| Quality Control: | QAM 20200414 |
| RNAse testing: | RNAses free 20200414 |
For fresh samples, only!!! Do not use VTM- frozen samples!!!
Protocol – Isolation of RNA from nasopharyngeal, buccal swabs
Comparison of the virus RNA detection from different matrices
Fresh (1) vs. Frozen (2) virus transfer media using ViRNAExTM RNA-extraction protocol
Compared viral RNA isolation protocols (magnetic beads-based method, column-based method) to ViRNAExTM extraction protocol in fresh and frozen virus transfer media of patients tested previously as COV2- positive. Obtained RNA has been used for GeneFirst™ qPCR testing of the genes: SARS-CoV-2- Gene N (A), SARS –COV2- ORF1ab (B), and epithelial cell control gene (C).
Analyzing the data from qPCR:
Figure 1: ViRNAExTM has comparable RNA-virus detection capacity if fresh virus transfer media is used for RNA isolation. The differences in Ct values were described on 24 samples and represent difference of
A. 1,7(Ct- cycle) for SARS –COV2-N gene)
B. 1,6 (Ct-cycle) for SARS-COV2-OFR1ab gene and
C. 3,3 (Ct- cycle) for epithelial control gene. Please see the Figure 1 A, B, C. The green amplification curve represents ViRNAExTM sample, the red one represents RNA separated by magnetic beads.
Figure 2: Similarly, on the Figure 2 the ViRNAExTM (Green , Red curve) is compared to the column-based RNA isolation (blue curve). In this case frozen virus transfer media of previously positive patients were used for comparisons (12 samples in total). We may see the significant delays in SARS-COV2- N gene (8 Ct cycles difference) and SARS- COV2 – ORF1ab gene (10 Ct cycles difference). The storage under freezing conditions did not influence the epithelial control gene detection. Green and red curves represent different starting concentrations of viral load.
CoVirED – Corona Virus Extraction and Detection kit
Product is composed from:
SARS-CoV-2 RT-PCR Detection Kit + ViRNAEx RNA Extraction Kit
Kit size: 96 tests
Kit includes: Primers and Probes, Reverse Transcriptase, DNA Polymerase and Positive Control and ViRNAEx RNA Extraction Kit
