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Development of a method for quantitative assessment of TREC and KREC molecules in whole blood and dried blood spots using digital PCR
https://doi.org/10.25557/2073-7998.2024.10.55-62
Abstract
A reduction in the indices of T- and B-lymphocytes is a common feature of a number of pathological conditions. The assessment of these indices is of paramount importance in the context of newborns, as the absence of timely diagnosis and treatment can result in the development of severe infectious diseases, disability, and high mortality. The objective of this study was to develop a highly sensitive and precise method for enumerating naïve T- and B-lymphocytes based on the quantification of excision circular DNA molecules TREC and KREC through digital polymerase chain reaction. Consequently, a specific set of primers and probes was developed, enabling highly accurate determination of the number of TREC and KREC molecules and counting of the number of nucleated cells. The proposed method exhibits high diagnostic specificity and sensitivity, rendering it suitable for use in both screening diagnostics of immunodeficiency states in newborns and for assessing the immune status in adults with various immunopathological conditions.
Supporting agencies: The study was carried out according to scientific research 123041700028-8 dated 04/17/2023.
About the Authors
A. A. Sleptсov
Tomsk National Research Medical Center of the Russian Academy of Sciences, Research Institute of Medical Genetics
Russian Federation
Russian Federation
10, Naberejnaya Ushaiki, Tomsk, 634050
D. S. Orlov
Tomsk National Research Medical Center of the Russian Academy of Sciences, Research Institute of Medical Genetics
Russian Federation
Russian Federation
10, Naberejnaya Ushaiki, Tomsk, 634050
S. A. Ogorodnikov
Tomsk National Research Medical Center of the Russian Academy of Sciences, Research Institute of Medical Genetics
Russian Federation
Russian Federation
10, Naberejnaya Ushaiki, Tomsk, 634050
M. S. Nazarenko
Tomsk National Research Medical Center of the Russian Academy of Sciences, Research Institute of Medical Genetics; Siberian State Medical University
Russian Federation
Russian Federation
10, Naberejnaya Ushaiki, Tomsk, 634050
2, Moskovsky trakt, Tomsk, 634050
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Review
For citations:
Sleptсov A.A., Orlov D.S., Ogorodnikov S.A., Nazarenko M.S. Development of a method for quantitative assessment of TREC and KREC molecules in whole blood and dried blood spots using digital PCR. Medical Genetics. 2024;23(10):55-62. (In Russ.) https://doi.org/10.25557/2073-7998.2024.10.55-62
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