Abstract
Russian real-time PCR (Q-PCR) kits and reference foreign kits were experimentally compared using serial dilutions (30–300,000 copies) of a plasmid containing a cDNA fragment of fusion PML-RARα (DNA standard). The Russian RialitiTM kit and ABI TaqMan PCR core reagent kit were similar in the efficiency of amplification (1.919 and 1.927, respectively) and in the coefficient of correlation obtained for the standard calibration plots (0.999 and 0.996, respectively). The RialitiTM kit was used to estimate the copy number of PML-RARα and ABL (internal control) in cDNA specimens obtained from peripheral blood and bone marrow of patients with acute promyelocytic leukemia and the chromosome translocation t(15;17), which generates chimeric PML-RARα. The PML-RARα copy number per μg total RNA ranged from 58,600 to 831,500 before therapy. After 3.5 months of chemotherapy, the patient showed no signs of the minimal residual disease: PML-RARα was undetectable by Q-PCR, which agreed with clinical data. Studies with the example of PML-RARα demonstrated the suitability of the RialitiTM kit for oncodiagnosis.
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Original Russian Text © O.Yu. Manzeniuk, S.G. Malakho, V.M. Pekhov, I.S. Kosorukova, A.B. Poltaraus, 2006, published in Molekulyarnaya Biologiya, 2006, Vol. 40, No. 2, pp. 349–356.
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Manzeniuk, O.Y., Malakho, S.G., Pekhov, V.M. et al. Characterization of Russian universal kits for real-time PCR: Application to molecular oncodiagnosis. Mol Biol 40, 305–311 (2006). https://doi.org/10.1134/S0026893306020178
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DOI: https://doi.org/10.1134/S0026893306020178