Development and laboratory testing of a prototype automated device for tuberculosis diagnosis using real-time PCR

This article presents the results of the development and laboratory testing of portable automated device prototype for rapid tuberculosis diagnosis and detection of drug resistance. The system comprises an analyzer and a disposable cartridge that contains all necessary reagents for sample preparation and PCR amplification inside. The analytical protocol includes sputum processing (ultrasonic treatment, lysis, and DNA extraction using magnetic particles), followed by nested real-time PCR for the detection of Mycobacterium tuberculosis and probe-melting analysis of rpoB gene mutations. Laboratory tests using sputum samples demonstrated that the system reliably detects M. tuberculosis DNA (IS6110 target) with high sensitivity, including samples with low bacterial load, and accurately identifies mutations associated with rifampicin resistance. An internal control confirms the absence of PCR inhibition. The accuracy of the results is comparable to standard laboratory PCR testing. Technological advantages of the system include full automation from sample to result (total analysis time ~2 hours) and reduced risk of contamination due to the sealed cartridge format compared to conventional molecular genetic methods. The developed platform demonstrates promise for point-of-care tuberculosis diagnostics, including multidrug-resistant forms. Further development is to expand the test panel to include resistance detection not only to rifampicin but also to isoniazid and fluoroquinolones.