DNA Extraction in TrueNAT MTB and TrueNAT MTB/RIF DX Assay

Equipment and Consumables

• Trueprep AUTO v2 Universal Cartridge Based Sample Prep Device
• Trueprep AUTO v2 Universal Cartridge Based Sample Prep Kit

Steps in DNA extraction (Molbio Trueprep Auto Universal Cartridge Based Sample Prep Device)

1. Wear gloves, mask.

2. Insert the plug-in connector to attach the reagent pack to the Trueprep device (Figure 1).

Figure 1: Trueprep Cartridge Based Sample Prep Kit attached to the Trueprep Auto Device using the Plug-in Connector; Source: Practical Guide to Implementation of Truenat Tests for the Detection of TB and Rifampicin Resistance

3. Disinfect the work surfaces with the freshly prepared 1% bleach, followed by 70% alcohol.

4. Open the cartridge pouch, take out the cartridge.

5. Label the cartridge (Patient ID, Date), place it on the cartridge stand.

6. Open the black cap of the cartridge sample chamber.

7. Using a pipette transfer 3 ml contents of the lysis buffer bottle (processing step) in the sample chamber.

8. Discard the pipette and lysis buffer bottle (1% sodium hypochlorite).

9. Re-cap the sample chamber.

10. Press the “Power” button to switch on the Trueprep device.

11. Press “Eject” to eject the cartridge holder (Figure 2), pull out gently.

Figure 2: Top view of the Trueprep AUTO Universal Cartridge-based Sample Prep Device; Source: Practical Guide to Implementation of Truenat Tests for the Detection of TB and Rifampicin Resistance

12. Insert the cartridge in the cartridge holder (sample chamber to the right when seen from the front).

13. Push to close the cartridge holder (the click sound confirms the correct cartridge loading).

14. Press the “Start” button (Figure 2).

15. DNA extraction begins, reagents from bottles are automatically added to the cartridge (pre-programmed) (Figure 2).

16. On completion (18-20 min), the cartridge holder automatically ejects.

17. Remove the cartridge, place it on the cartridge stand.

18. Label the ECT tube (Elute Collection Tube) with Patient ID & Date.

19. Pierce the covering of elute compartment in the cartridge with a filter barrier pipette tip, aspirate elute (DNA) into the ECT tube.

20. Discard the used pipette tips, used cartridge (1% sodium hypochlorite).

21. DNA can be used immediately for amplification/ stored in fridge (4°C for up to 24 hours) or -20°C (for up to 1 year).

Resources

• Practical Guide to Implementation of TrueNAT Tests for the Detection of TB and Rifampicin Resistance.
• Operation of Molbio Trueprep AUTO Universal Sample Prep Device.
• Molbio Trueprep Auto Universal Cartridge-based Sample Prep Device.

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