Swabs, immediately after collection, should be placed into sterile collection tubes containing aliquoted PCR DIRECT ETM.
We recommend preparing sample collection tubes ahead with 300 µl of PCR DIRECT ETM per tube.
For maximum sensitivity in direct RT-qPCR PCR, the reaction may be prepared with up to 50% sample by volume (e.g., up to 12.5 µl sample in a 25 µl reaction).
If a bead- or column-based isolation kit will be used, 200-300 µl of sample in PCR DIRECT ETM may be used for subsequent processing.
- Collect oral or nasal swabs according to laboratory procedure.
- Place swab head into PCR DIRECT ETM; swirl and rotate swab head against the tube wall 10 times to extract the maximum amount of liquid from the swab head.
- Begin to remove the swab, press the swab head against the side of the collection tube to maximize the amount of prepared lysate.
- Shake the tube vigorously at least ten times.
- A sample is ready for transport and subsequent downstream molecular analysis or processing.
Note: the head of the swab should be fully submerged in a minimal volume of PCR DIRECT ETM. Complete immersion will ensure that the entire collected sample has been lysed. Minimizing the volume of PCR DIRECT ETM ensures the highest concentration of nucleic acids and supports direct PCR testing methods.
For saliva samples, donors should be instructed not to consume food or drink, or use gum, toothpaste for at least 30 minutes prior to sample collection.
Saliva should not be mixed with other transport media or lysis reagents prior to the addition of the PCR DIRECT ETM as this may affect results.
This protocol may be adjusted for larger sample volumes by scaling the volume of PCR DIRECT ETM in proportion to the increase in the sample volume.
If a bead- or column-based isolation kit will be used, 200-400µl of sample in PCR DIRECT ETM may be used for subsequent processing.
- Add 100 µl of saliva to a 1.5 ml low binding microtube.
- Add 235 µl of PCR DIRECT ETM to the sample with the same pipette, mix ten (10) times.
- Sample is ready for transport and subsequent downstream molecular analysis or processing.
Note: Inadequate mixing in step 2 will result in lower sensitivity. Mixing by pipetting is preferred.