Q: Can this kit be used as rapid-test?
No. This kit can only be used in fully equiped laboratories which have a RealTime PCR reader.
Q: Is the kit compatible with any Reader?
The kit is compatible with any Real-Time PCR Device able to detect FAM and HEX (and ROX, if required).
Q: Which specimens can be examined?
Respiratory specimens including nasopharyngeal / oropharyngeal aspirates or washes, nasopharyngeal oropharyngeal swabs, broncheoalveolar lavage, tracheal aspirates and sputum. Swab specimen s should be collected only on swabs with a synthetic tip (such as polyester or Dacron®) with aluminum or plastic shafts. Swabs with calcium alginate or cotton tips with wooden shafts are not acceptable.
Q: Additional equipment needed?
- Suitable means & equipment for nucleic acid extraction
- Real-time PCR detection system equipped for FAM™ detection
- Adjustable pipettes & fitting filtered pipette tips
- Appropriate PSA & workspaces for working with potentially infectious samples
- Surface decontaminants such as DNAZap™ (Life Technologies), DNA Away™ (Fisher Scientific), RNAse Away™ (Fisher Scientific), 10 % bleach (1:10 dilution of commercial 5.25-6.0 % sodium hypochlorite)
- Nuclease-free tubes / strips / plates to prepare dilutions, mastermixes etc.
- Nuclease-free tubes / strips / plates corresponding to the PCR device
- Suitable storage options for reagents and specimen (4 °C, -20 °C, -70 °C)
Q: What if dH2O controls (NTC) give a positive Ct for any assay?
If they do, the reaction was contaminated with sample RNA / DNA. Decontaminate equipment and workspace and repeat the reactions.
Q: If the HEC fails to amplify, must the sample still be considered positive?
For a sample to be considered positive for 2019 nCoV , both assays (E / RdRP) must give positive Ct values. If the HEC fails to amplify, the sample must still be considered positive.
Q: When is a sample to be considered negative for 2019-nCoV?
For a sample to be considered negative for 2019 nCoV , none of the 2 assays (E / RdRP) must give positive Ct. The HEC must give a positive Ct value (< 35 cycles) for these samples to ensure that sample material of suitable quality was present.
Q: What if Ct values for HEC are >35 cycles?
All reactions containing RNA isolate must give positive Ct values for the HEC assay. The Ct values should be < 35 cycles. Failure to amplify the negative human extraction control indicates a flawed RNA extraction or loss of RNA isolate due to RNAse contamination. The sample is not sufficient, results cannot be interpreted.
Q: What if negative Ct values for TPC are observed?
When using the TPC for 2019 nCoV , a positive Ct for both assays must be observed . The Ct value for the TPC should be < 35 cycles. If the Ct value does not correspond to the expected value or not all assays are tested positive , PCR was compromised . Check the reaction setup and PCR device settings and repeat the reactions. Repeated freeze and thaw cycles of the TPC can compromise its quality resulting in late Ct values.
Q: What if no amplification signal can be observed?
If no amplification signal is observed for any assay, PCR was inhibited. Check reaction setup and device settings and repeat the RNA extraction if necessary. Results are invalid and cannot be interpreted.
Q: What if only the E gene assay is observed to be positive?
If < 2 of the target assays are positive (e.g. only E gene, etc.), results are inconclusive. Check reaction setup and device settings and repeat the RNA extraction if necessary. Results are invalid and cannot be interpreted.
Q: I do not see a price for this kit. Instead it says "Contact our Customer Service for availability and price in your country."
Due to our supplier aggreement we are not allowed to sell this kit in Denmark, Turkey and UK. If you are located in another country please contact us and we will issue a quote for this kit.