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).

Inbuilt Controls used in TrueNAT 

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)).

 

Truenat Cartridge and Inside Demonstration

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:

Fluorescence Microscopy Using LED Microscope

Fluorescence Microscopy is a newer and better type of microscopy where the TB bacteria are stained using a fluorescent dye using the property of Acid Fastness. The dye will fluoresce when illuminated by UV light. When the UV light source is an LED Lamp, it is called LED fluorescent microscopy.

Here the bacilli appear as slender bright yellow fluorescent rods, standing out clearly against a dark background, as can be seen in the figure below.

Requesting a Test on Nikshay

Once a presumptive TB patient is identified, the patient is enrolled online by a healthcare worker or doctor in Nikshay online portal. For diagnosis of Tuberculosis, the treating physician can request a test utilizing the request test option of Nikshay online portal. The step-by-step approach for test request is as follows:

 

Step 0: Go to the Patients Page.

Step 1: Select the “Tests” tab.

Step 2: Click the “Add Test” button.

 

Specimen Carriers

Technical specifications for TB diagnosis specimen carriers are as follows:

 

Thermocol Box:

  • Thickness of box - 2.5 cm
  • Outer dimensions: Length - 18.5 cm, breadth - 13 cm, height - 12 cm (without lid), height - 14 cm (with lid); Inner dimensions: Length - 14.5 cm, breadth - 8 cm, height - 12 cm (without lid), height - 13 cm (with inner part of lid).

 

Gel Pack:

Overview of the Patient Management Workflow in Nikshay

Nikshay is an integrated Information, Communication and Technology (ICT) platform adopted by the National Tuberculosis (TB) Elimination Programme (NTEP) for TB patient management and care.

 

The patient management workflow in Nikshay streamlines a series of activities required for TB patients from identification of presumptive TB cases to recording treatment outcomes. Nikshay uses in-built modules in the platform to perform these tasks. There are also different user logins that allow certain functions (see figure below).

 

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