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Accurate, Rapid, Robust and Economical diagnostic technoliogieS for Tuberculosis

Periodic Reporting for period 2 - ARREST-TB (Accurate, Rapid, Robust and Economical diagnostic technoliogieS for Tuberculosis)

Reporting period: 2020-07-01 to 2021-06-30

ARREST-TB is a paradigm changing, multilateral project focused on the development of frugal diagnostic technologies for tuberculosis. The team brings together a revolutionary DNA profiling technology and “no wash” optical labelling of patient sputum, with a focus and specific design of deployment in primary care and low resource settings. It addresses the key needs of patients, clinicians and the World Health Organisation's stated deliverables of “affordable, reliable and point-of-care TB diagnostics, with the vision of removing the need for centralised facilities and offering remote primary care diagnosis coupled with telemetry for “cloud based” notification to ensure public health and surveillance interventions.

The core technology deliverables will include:
• Rapid molecular profiling of drug resistance: Translating molecular diagnostics based on dynamic chemistry into a low-cost assay to allow the accurate and rapid screening for Mycobacterium tuberculosis and drug-resistance ‘in-field’, thus providing rapid screening of at risk groups for TB as well as identification of drug resistance profiles.
• Rapid screening of TB infection with the development of low-cost portable optical devices and chemical reagents for accurate and rapid ‘triage tests’ to allow the detection of Mycobacteria ‘in-field’ in sputum at costs lower than existing microscopy-based methods. This technology has the aim of replacing current sputum-based microscopy methods.
• Biomarkers for early diagnosis and assessing treatment response: We are developing point-of-care diagnostics using small blood samples.
These tests will allow rapid detection of TB/MDR-TB with the use of an ‘app’ on mobile-phones to interpret results, log details of the location, as well as transmitting the data and results to the cloud for collation and reporting. This will help reduce under-reporting of TB cases, help track treatment outcomes as well as identifying ‘hot spots’ of TB, thereby assisting in the determination of clusters of transmission to target public health interventions.
Thus, ARREST-TB aims to develop technologies that are economically viable in the context of resource poor settings – with the whole project driven by practical need and local resourcing constraints. The solutions require negligible initial costs for implementation, minimal training and expertise for operation, and significantly lower running costs than current detection technologies, thereby addressing a key objective of the Horizon 2020 work program in terms of ‘addressing the needs of the most vulnerable groups’.
- Frugal devices for TB and MDR TB diagnosis: We have developed novel low-cost assays, (based on DestiNA technology) that will allow accurate and rapid screening ‘in-field’ of TB and NTM infections, thus providing the ability for rapid screening of at risk groups for TB. These devices have undergone successful independent evaluation in the EU and Russia. Shortly, final evaluations / validations of these devices will be completed in India and Russia.

- DNA extraction device: The first prototype of a portable, battery-powered, low cost extraction device has been developed by HWU, with the intention of being used ‘in field’, providing Mycobacterial DNA of high quality suitable for further amplification and detection. Further optimisation is in progress to deliver the final device by the end of the project.

- Device for microRNA-based diagnosis (using patient serum) of latent TB, active TB, risk of progression from latent to active TB and monitoring treatment outcome. The devices are ready, and have been successfully evaluated preliminarily in the EU and ready for thorough evaluations in Russia and India. Biological samples for the biomarker discovery study have been obtained, and the study is due to begin shortly.

- Rapid triage using No wash’ chemical probes for staining mycobacterium and optical devices for detecting mycobacterium. We have developed low-cost portable optical devices and molecular probes for accurate and rapid ‘triage tests’ to allow rapid ‘no-wash’ detection of Mycobacterium tuberculosis complex ‘in-field’ in sputum at costs comparable with or lower than existing microscopy-based methods. Evaluations in progress in the UK, Italy, Russia and India – based on which modifications to the design, as well as staining protocols are being implemented. The first versions of the optical device has been fabricated and being improved.
Rapid ‘triage’ using molecular probes and low cost portable optical devices: Due to its low cost and long history of use, sputum smear microscopy is widely used to screen for suspected pulmonary TB. However, these microscopy tests depend on stain-based methods that were developed over 100 years ago and require extensive processing and washing steps, resulting in detection levels as low as ~30%, depending on factors such as experience of the user and quality of sample. Therefore, we have developed fluorescent compounds (under evaluation in India and Russia) that will specifically stain Mycobacteria, avoiding non-specific labelling of other Gram +ve and –ve bacteria and other components in the sputum, thereby providing a ‘no-wash’ visualisation of the bacteria that cause tuberculosis.

Rapid ‘in-field’ molecular diagnosis of TB using DestiNA technology: ARREST-TB is well on its way to deliver a low cost, and reliable nucleic acid detection platform for diagnosis of Mycobacterium tuberculosis complex, Non-tuberculosis Mycobacteria, as well as their drug-resistant mutations in the field. Up until now, such testing has required laborious, and time-consuming culture or expensive PCR based techniques that require specialised expertise and instrumentation, high installation, running and maintenance costs, cold-chains and so-forth, scarce in many primary care and low resource settings.

Non-sputum test for detecting TB with DestiNA technology using microRNAs as biomarkers: Biomarkers in easy-to-collect specimens such as saliva, urine, serum and whole blood have the potential to detect tuberculosis exposure and differentiate between active TB disease and latent TB infection and monitor treatment response. Despite significant investments in research over the past decades resulting in numerous reports on new biomarker candidates, a versatile biomarker-based test has not materialised for active TB, latent TB and to predict t the risk of progression from latent to active TB. Therefore, we are developing efficient and affordable microRNA-based assays using the Destina-Optoi platform.

Mobile phone ‘app’ for data interpretation, collection and remote collation: The aim here is to have the all the detection (and extraction) devices run by an ‘app’ on mobile phones. This ‘app’ will also enable telemetry, i.e. interpretation of results, logging of the details of the experiment and location (GPS), as well as transmitting the data and results to the cloud for collation and reporting. This will reduce under-reporting of TB cases, help track treatment outcomes as well as rapidly identifying ‘hot spots’ of TB.
ARREST-TB consortium during the 9-month meeting in Edinburgh