Clinical results for a new Russian reagent kit based on the polymerase chain reaction for qualitative and quantitative in vitro determination of tuberculosis DNA

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BACKGROUND: The global fight against tuberculosis (TB) continues. The timely prescription of anti-TB therapy requires the rapid and effective identification of the causative agent, the Mycobacterium tuberculosis complex. Molecular genetic testing is used to detect the pathogen’s DNA and to provide results on the day the biological material is submitted to a laboratory. The Center for Strategic Planning and Management of Biomedical Health Risks has developed an AmpliTest® MBT reagent kit for the qualitative and quantitative determination of M. tuberculosis complex DNA in samples of human biological material and bacterial cultures using polymerase chain reaction.

AIM: The aim of the study was to evaluate the efficacy of the new AmpliTest® MBT reagent kit compared to a similarly designed kit approved in Russia as an in vitro diagnostic medical device, based on pre-approval clinical studies.

MATERIALS AND METHODS: A total of 575 samples were evaluated, including 475 biological samples obtained from patients with pulmonary and extrapulmonary tuberculosis: sputum, n =115; bronchoalveolar lavage, n =109; biopsy (surgical material), n =145; urine, n =106; and 100 bacterial culture samples. All samples, except cultures, were processed with NALC-NaOH reagents to obtain precipitates, each of which was divided into four equal parts and analyzed using three versions of the AmpliTest® MBT kit and the reference AmpliTub-RV kit (Syntol LLC). Amplification was performed using CFX96, DTprime, Applied Biosystems QuantStudio 5, Rotor-Gene Q manually or with software to automatically record results. The efficacy parameters of the AmpliTest® MBT kit were calculated, i.e. positive and negative concordance of results (with 95% confidence interval). Linear correlation coefficient (R2) was determined for quantitative results.

RESULTS: There was complete concordance between the results of the qualitative determination of M. tuberculosis complex DNA in the samples tested using the new kit and the reference kit. The efficacy rates of the new AmpliTest® MBT kit were 100% for all samples. The linear correlation coefficients (R2) of the quantitative determinations of M. tuberculosis complex DNA were greater than 0.9.

CONCLUSION: Clinical studies of the new AmpliTest® MBT reagent kit demonstrated its high efficiency in the qualitative and quantitative determination of M. tuberculosis complex DNA in samples of human biological material and bacterial cultures compared to the reference kit.

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Sobre autores

Yuliya Mikulovich

Centre for Strategic Planning and Management of Biomedical Health Risks

Autor responsável pela correspondência
Email: YuMikulovich@cspfmba.ru
ORCID ID: 0009-0006-3662-7528
Código SPIN: 8676-9256

Cand. Sci. (Chemistry)
Rússia, 1 bldg., 10 Pogodinskaya street, 119121 Moscow

Anastasiya Zaytseva

Centre for Strategic Planning and Management of Biomedical Health Risks

Email: AZaytseva@cspfmba.ru
ORCID ID: 0009-0007-9297-0256
Rússia, 1 bldg., 10 Pogodinskaya street, 119121 Moscow

Sergei Lazebnyi

Moscow regional clinical antitubercular clinic

Email: nelazebnyi@mail.ru
ORCID ID: 0000-0002-8667-3614
Rússia, Mytishchi

Maria Plekhanova

Moscow regional clinical antitubercular clinic

Email: dina-plus@mail.ru
ORCID ID: 0000-0003-1687-7598
Código SPIN: 3595-8608

MD, Dr. Sci. (Medicine)
Rússia, Mytishchi

Sergei Smerdin

Moscow regional clinical antitubercular clinic

Email: mz_mokptd@mosreg.ru
ORCID ID: 0000-0002-2602-214X
Código SPIN: 7649-0300

MD, Dr. Sci. (Medicine), Professor
Rússia, Mytishchi

German Shipulin

Centre for Strategic Planning and Management of Biomedical Health Risks

Email: Shipulin@cspfmba.ru
ORCID ID: 0000-0002-3668-6601
Código SPIN: 1908-9098

MD, Cand. Sci. (Medicine)
Rússia, 1 bldg., 10 Pogodinskaya street, 119121 Moscow

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2. Fig. 1. Correlation graph of MTB DNA quantitative results in sputum samples obtained using the AmpliTest® MTB kit form 1 and the comparison Amplitub-RT kit.

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3. Fig. 2. Correlation graph of MTB DNA quantitative results in bronchoalveolar lavage samples obtained using the AmpliTest® MTB kit form 1 and the comparison Amplitub-RT kit.

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4. Fig. 3. Correlation graph of MTB DNA quantitative results in biopsy (surgical material) samples obtained using the AmpliTest® MTB kit form 1 and the comparison Amplitub-RT kit.

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5. Fig. 4. Correlation graph of MTB DNA quantitative results in urine samples obtained using the AmpliTest® MTB kit form 1 and the comparison Amplitub-RT kit.

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6. Fig. 5. Correlation graph of MTB DNA quantitative results in bacterial culture samples obtained using the AmpliTest® MTB kit form 1 and the comparison Amplitub-RT kit.

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Declaração de direitos autorais © Mikulovich Y.L., Zaytseva A.I., Lazebnyi S.V., Plekhanova M.A., Smerdin S.V., Shipulin G.A., 2025

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