Report

Teaser, summary, work performed and final results

Periodic Reporting for period 1 - PITBUL (Point-of-Care implementation of TB testing with ultra-fast Local Heating PCR)

Teaser

The aim of this project is to research and develop the ultrafast Local Heating Polymerase Chain Reaction (LPCR) technology and its laboratory demonstrator from a non-integrated into an integrated system, for clinical proof-of-concept testing in tuberculosis (TB) centers in...

Summary

The aim of this project is to research and develop the ultrafast Local Heating Polymerase Chain Reaction (LPCR) technology and its laboratory demonstrator from a non-integrated into an integrated system, for clinical proof-of-concept testing in tuberculosis (TB) centers in Italy, Latvia, and Tanzania. This will imply a fully automated workflow in the demonstrator, all necessary reagents stored onboard of the prototype cartridge, and a cloud-based analysis software. An ultrafast point-of-care test (POCT) for TB and emerging TB multi-drug resistant markers (MDR) will allow patients with active TB to be rapidly and accurately diagnosed and adequately treated.

Work performed

At the project’s kick-off meeting, specifications for the POCT system and workflow were firstly defined by all partners, in particular the technical and user requirements based on ease of use, turn-around-time, sensitivity, specificity, safety, and costs. This established the basis for a development plan of the complete workflow, namely sample processing, instrument, cartridge and software.
In favor of the system’s portability, cost of goods, and sample purification abilities, the project consortium made the decision early on to focus exclusively on the Local Heating PCR (LPCR) variant that can be employed in a compact and portable system.
A complete manual workflow concept from sample to result was successfully demonstrated, establishing: an easy-to-use and user-safe sputum liquefaction, Mycobacteria lysis protocols, sample purification, and a TB assay with LPCR. This workflow was first demonstrated at GNA using surrogate sputum and manual pipetting on a prototype chip. The same was later successfully demonstrated by partners with TB-negative patient sputum spiked with TB from culture. This served as a proof of concept for the subsequent integration of the separate steps into one single, disposable cartridge, namely: lysis, target DNA capture plus sample purification, and LPCR (amplification and detection).

Final results

The workflow concept can allow for TB testing near the patient at an unprecedented speed. A safe sample handling and transfer process have been designed, beyond the current state of art. Furthermore, assays for highly relevant, but largely neglected mutations responsible for drug-resistance in TB will be designed and tested. These aspects can contribute to a broad implementation of the future test system.

Website & more info

More info: https://www.pitbul-project.eu.