Achieve high sensitivity using dry swab without isolation kits

  • Simplify workflow – reduce consumables and kits, and eliminate digestion step

  • Deliver sensitivity equivalent to CDC protocol without capture and concentrate

  • Mucolytic properties reduce lab failures or repeats

  • Mitigate reagent supply issues with a single reagent

  • Avoid transport media to reduce burden of validation and inconsistency across suppliers

  • Hazard-free chemistry assures safety with minimal cost

To realise the advantages of Swab Extraction Reagent plus add viral inactivation, stabilisation and ambient transport at point of collection, see alternative PCR DIRECT Extracting Transport Medium

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Need

The use of buffers or standard transport media with swabs can be problematic:

  • Media dilute nucleic acids
  • Requires capture and concentrate steps and associated costs to achieve desired sensitivity
  • Incoming samples treated with transport media from multiple suppliers creates challenges for lab standardisation
  • Isolation kits contain hazardous chemicals

Solution

Simplify collection and sample prep by treating dry swab with  Swab Extraction Reagent

  • Extracts nucleic acids from dry swabs
  • Delivers sensitivity equivalent to CDC protocol without capture and concentrate or digestion step – saves costs of time, kits and consumables
  • Avoid validation challenges of receiving samples in multiple transport media
  • Guanidine-free formulation eliminates safety risks and reduces waste disposal costs

Standard 300µl Swab Protocol

  1. Collect the dry swab according to laboratory procedure, in a dry tube without transport media.
  2. Add 300µl of PCR DIRECT® ER1 to a low binding micro tube (1.5-2.0ml)
  3. Add the swab to the micro tube, squeeze and turn the swab against the micro tube wall to extract as much as possible of the mucus, at least ten (10) times
  4. Pipette mix at least ten (10) times
  5. Heat sample for five (5) minutes at 70°C (Optional)
  6. Sample is ready for downstream molecular analysis or processing

Note: Inadequate mixing,  squeeze and turns in steps 3 and 4 will result in lower sensitivity. Mixing by pipetting is preferred

Nucleic acid yield and processing efficiency may vary depending on the virus extracted. Samples prepared with the PCR DIRECT ER1 should be used in the intended downstream processes within 4 hours of preparation or frozen, if intended for use within 24 hours. Freeze-thaw cycles after adding PCR DIRECT ER1 should be limited to 2 or fewer. For longer term storage of RNA, we recommend the addition of an RNase inhibitor in the amount recommended by the manufacturer to ensure long term stability.

High Sensitivity SARS-CoV-2 Testing

PCR DIRECT Swab Extraction Reagents were used to extract viral RNA from contrived clinical swab samples; NP swab samples were collected from healthy donors and spiked with heat-inactivated SARS-CoV-2 virus.

Limit of Detection - Swab Extraction Reagent

Swab samples were processed with PCR DIRECT Swab Extraction Reagents and tested via qRT-PCR (CDC N1 assay). Reliable, consistent, detection was observed at 900 viral copies per swab, which indicate that PCR DIRECT Swab Extraction Reagents can form part of high sensitivity testing workflows.

As a control, human RNA was also detected in the same swab samples using the CDC human RNaseP assay.

All results generated using Thermo Fisher Taqman Fast Virus 1-Step Master Mix.

 

Our PCR DIRECT Swab Extraction Reagent:

  • Simplifies and accelerates the processing of dry swabs, with non-hazardous chemicals.
  • Are automatable and scalable and can be used in high throughput sample processing labs.
  • Suitable for a range of swab types including flocked swab or NP-OP swabs.

Product code

UFL141, UFL145

Sample type

Nasopharyngeal, Oropharyngeal, Nasal Swab

Sample input volume

One dry swab

Reaction time

1 minute hands on

No. of reactions

40, 400

Storage

Store at room temperature, do not freeze

Shipping conditions

Room temperature