Multiplex droplet digital PCR for 22q11.2 microdeletions screening and DiGeorge syndrome diagnostics.

Autor: Oscorbin IP; Laboratory of Pharmacogenomics, The Institute of Chemical Biology and Fundamental Medicine of the Siberian Branch of the Russian Academy of Sciences (ICBFM SB RAS), Novosibirsk 630090, Russia. Electronic address: osc.igor@gmail.com., Gordukova MA; G.N. Speransky Children's Hospital No. 9, Moscow, Russia., Davydova NV; G.N. Speransky Children's Hospital No. 9, Moscow, Russia., Zinovieva NV; G.N. Speransky Children's Hospital No. 9, Moscow, Russia., Kovzel EF; Clinical Immunology, Allergology, Pulmonology Program, Corporate Fund 'University Medical Center' of Nazarbayev University, Astana, Kazakhstan., Andries L; Laboratory of Clinical Immunology and Allergology, Nicolae Testemitanu State University of Medicine and Pharmacy of the Republic of Moldova, Chișinău, Moldova., Kudlay DA; The Department of Pharmacology, Faculty of Medicine, I.M. Sechenov First Moscow State Medical University, Pogodinskaya St. 1, Moscow 119991, Russia., Filipenko ML; Laboratory of Pharmacogenomics, The Institute of Chemical Biology and Fundamental Medicine of the Siberian Branch of the Russian Academy of Sciences (ICBFM SB RAS), Novosibirsk 630090, Russia.
Jazyk: angličtina
Zdroj: Clinica chimica acta; international journal of clinical chemistry [Clin Chim Acta] 2024 Sep 15; Vol. 563, pp. 119903. Date of Electronic Publication: 2024 Aug 08.
DOI: 10.1016/j.cca.2024.119903
Abstrakt: Background and Aims: DiGeorge syndrome (DGS) is a genetic disorder manifesting in polymorphic symptoms related to developmental abnormalities of various organs including thymus. DGS is caused by microdeletions in the 22q11.2 region between several low copy repeats (LCR) occurring in approximately 1 in 4000 live births. Diagnosis of DGS relies on phenotypic examination, qPCR, ultrasound, FISH, MLPA and NGS which can be relatively inaccurate, time-consuming, and costly.
Materials and Methods: A novel multiplex droplet digital PCR (ddPCR) assay was designed, optimized and validated for detection and mapping 22q11.2 microdeletions by simultaneous amplification of three targets - TUPLE1, ZNF74, D22S936 - within the deletion areas and one reference target - RPP30 - as an internal control.
Results: The assay reliable identified microdeletions when the template concentration was >32 copies per reaction and successfully detected LCR22A-B, LCR22A-C, LCR22A-D, and LCR22B-C deletions in clinical samples from 153 patients with signs of immunodeficiency. In patients with the microdeletions, flow cytometry detected a significant increase in B-cell and natural killer cell counts and percentages, while T-cell percentages and T-cell receptor excision circle (TREC) numbers decreased.
Conclusion: The designed ddPCR assay is suitable for diagnosing DGS using whole blood and blood spots.
Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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