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  • Truenat Cartridge and Inside Demonstration

    Content

    Truenat cartridges are components of Trueprep AUTO v2 Universal Cartridge Based Sample Prep Kit. As shown in the figure below, each cartridge is:

    • Used with Trueprep Device for DNA extraction and purification
    • Disposable
    • Single-use
    • Preloaded with Internal Positive Control.

    Truenat cartridges are used with Trueprep devices for DNA extraction and purification. The components of the cartridge are illustrated in the following figure and include:

    • Sample Chamber - Processed samples (pretreated with lysis buffer) are added to the sample chamber of the cartridge for processing on the Trueprep device.
    • Matrix Chamber - DNA released by chemical and thermal lysis cells binds to the proprietary matrix here.
    • Elute Chamber - Trapped DNA is washed with buffers to remove Polymerase Chain Reaction (PCR) inhibitors and is eluted from the matrix using the elution buffer. The purified DNA is collected from here.
    • Waste generated is contained within the dump region inside the cartridge.

    Figure: Inside of a TrueNAT Cartridge; Source: Practical Guide to Implementation of TrueNAT Tests for the Detection of TB and Rifampicin Resistance

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  • Consumables Required at a Truenat Lab

    Content

    Consumables required for Truenat test provided by the manufacturer include:

     

    Trueprep AUTO MTB Sample Pre-treatment Pack for sample processing

    1. Liquefaction buffer
    2. Lysis buffer
    3. Disposable transfer pipette (graduated) - 1ml

    Trueprep AUTO v2 Universal Cartridge Based Sample Prep Kit for DNA extraction

    1. The Reagent Pack contains the following reagents
      1. Wash Buffer A
      2. Wash Buffer B
      3. Elution Buffer
      4. Priming Waste
    2. The Cartridge Pack contains the following
      • Cartridge
      1. Elute collection tube
      2. Elute collection tube label
      3. Disposable transfer pipette
    3. Disposable Transfer Pipettes (graduated) - 3 ml
    4. Reagent Reset Card

    Truenat MTB Chip-based Real-Time PCR test for Mycobacterium tuberculosis

    1. Truenat MTB micro-PCR chip
    2. Microtube with freeze-dried PCR reagents
    3. DNase & RNase free pipette tip

    Truenat MTB-RIF Dx Chip-based Real-Time PCR Test for Rifampicin Resistant Mycobacterium tuberculosis

    1. Truenat MTB-RIF Dx micro-PCR chip
    2. Microtube with freeze-dried PCR reagents
    3. DNase & RNase free pipette tip

    Other Consumables

    1. Gloves
    2. Masks
    3. Sodium hypochlorite
    		  

     

       

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      1. Overview of the Truenat Testing Process

        Content

        The Truenat Assay Technology works on Real-time Polymerase Chain Reaction (PCR).

         

        The key steps include:

        1. Collection of specimens from a presumptive TB/known TB Patient (for UDST).
        2. Liquefaction and lysis of specimen using the Trueprep AUTO MTB sample pre-treatment pack (20 minutes)
        3. Extraction and purification of DNA using Trueprep AUTO v2 Universal Cartridge-based sample prep kit and Trueprep AUTO v2 Universal Cartridge-based sample prep device (20 minutes)
        4. Amplification of extracted DNA by the Truelab Real-time micro-PCR analyzer using freeze dry PCR reagent in microtubes on a Truenat MTB chip (35 minutes)
        5. DNA from positive test results is tested using Truenat MTB RIF Dx chip as a reflex test (55 minutes)

         

        The process flow from sample to result for Truenat Assay Technology is described in the figure below.

         

        Figure: Process flow from sample to result for Truenat Assay Technology; Source: Practical Guide to Implementation of Truenat™ Tests for the Detection of TB and Rifampicin Resistance.

         

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      2. Truenat MTB and RIF Assay as a TB Diagnostic Test

        Content

         

        The tests are performed using Trueprep AUTO Sample Pre-treatment and Prep kits and Truenat micro-PCR chips.

         

        To detect M. tuberculosis, the Truenat MTB chip amplifies a portion of the ribonucleoside-diphosphate reductase gene, nrdB with a Limit of Detection (LOD) of about 100 Colony Forming Units (CFU)/ml sputum sample.

         

        DNA extraction and detection of M. tuberculosis takes approximately one hour.

         

        When M. tuberculosis is detected, as a follow-on test, a small volume of the already extracted DNA is used to test for resistance to Rif using Truenat MTB Rif Dx chip.

         

        Mutations associated with Rif-resistance within the RRDR region of rpoB gene are detected using a probe melt assay and read using the Truelab micro-PCR Analyzer.

         

        The detection of Rif-resistance takes an additional one hour.

         

        Truenat Testing Capacity

        • The Truelab Analyzer is available as 1 (Uno), 2 (Duo) or 4 (Quattro) chip ports (figure below).
        • The different ports can be used independently to test multiple samples.
        • In an eight-hour daily work shift, the estimated throughput of Truelab Analyzer Uno, Duo or Quattro can be 7-9, 15-18, 30-36 specimens, respectively.

         

        Results Reporting 

        A Truelab micro PCR printer (figure below) is used to print test results. It can also be connected via SIM card/ Wi-Fi/ Bluetooth to transmit results.

         

        Figure: Different Equipments used in the Truenat Workstation

         

         

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      3. Inbuilt Controls used in TrueNAT 

        Content

        Truenat tests use Internal Quality Control to assess test validity using Internal Positive Control (IPC).

         

        • The cartridge contains pre-loaded IPC.
        • IPC is part of the full process control.
        • IPC undergoes all processing steps along with specimen from extraction to amplification.
        • IPC assesses the validity of the test run from sample processing to result and is illustrated in following figures (Figure 1(A) and Figure 1(B)).

         

        Figure 1(A): Amplification of IPC in Valid Test; Source: MolBio Diagnostics Pvt. Ltd.

         

         

        Figure 1(B): Invalid Test with Failure of IPC Amplification; Source: MolBio Diagnostics Pvt. Ltd.

         

         

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