SMIRNOVA, Anastasia O, Anna M MIROSHNICHENKOVA, Yulia V OLSHANSKAYA, Michael A MASCHAN, Yuri B LEBEDEV, Dmitriy CHUDAKOV, Ilgar Z MAMEDOV a Alexander KOMKOV. The use of non-functional clonotypes as a natural calibrator for quantitative bias correction in adaptive immune receptor repertoire profiling. elife. CAMBRIDGE: ELIFE SCIENCES PUBLICATIONS LTD, 2023, roč. 12, Jan, s. 1-17. ISSN 2050-084X. Dostupné z: https://dx.doi.org/10.7554/eLife.69157.
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Základní údaje
Originální název The use of non-functional clonotypes as a natural calibrator for quantitative bias correction in adaptive immune receptor repertoire profiling
Autoři SMIRNOVA, Anastasia O, Anna M MIROSHNICHENKOVA, Yulia V OLSHANSKAYA, Michael A MASCHAN, Yuri B LEBEDEV, Dmitriy CHUDAKOV (643 Rusko, domácí), Ilgar Z MAMEDOV a Alexander KOMKOV (garant).
Vydání elife, CAMBRIDGE, ELIFE SCIENCES PUBLICATIONS LTD, 2023, 2050-084X.
Další údaje
Originální jazyk angličtina
Typ výsledku Článek v odborném periodiku
Obor 30102 Immunology
Stát vydavatele Velká Británie a Severní Irsko
Utajení není předmětem státního či obchodního tajemství
WWW URL
Impakt faktor Impact factor: 7.700 v roce 2022
Kód RIV RIV/00216224:14740/23:00133611
Organizační jednotka Středoevropský technologický institut
Doi http://dx.doi.org/10.7554/eLife.69157
UT WoS 000931254200001
Klíčová slova anglicky immune receptor repertoire; high-throughput sequencing; multiplex PCR; PCR bias correction; adaptive immunity; TCR
Štítky rivok
Příznaky Mezinárodní význam, Recenzováno
Změnil Změnila: Mgr. Eva Dubská, učo 77638. Změněno: 8. 3. 2024 10:10.
Anotace
High-throughput sequencing of adaptive immune receptor repertoires is a valuable tool for receiving insights in adaptive immunity studies. Several powerful TCR/BCR repertoire reconstruction and analysis methods have been developed in the past decade. However, detecting and correcting the discrepancy between real and experimentally observed lymphocyte clone frequencies are still challenging. Here, we discovered a hallmark anomaly in the ratio between read count and clone count -based frequencies of non-functional clonotypes in multiplex PCR- based immune repertoires. Calculating this anomaly, we formulated a quantitative measure of V-and J -genes frequency bias driven by multiplex PCR during library preparation called Over Amplification Rate (OAR). Based on the OAR concept, we developed an original software for multiplex PCR-specific bias evaluation and correction named iROAR: immune Repertoire Over Amplification Removal (https://github.com/smiranast/iROAR). The iROAR algorithm was successfully tested on previously published TCR repertoires obtained using both 5' RACE (Rapid Amplification of cDNA Ends) -based and multiplex PCR- based approaches and compared with a biological spike -in -based method for PCR bias evaluation. The developed approach can increase the accuracy and consistency of repertoires reconstructed by different methods making them more applicable for comparative analysis.
VytisknoutZobrazeno: 1. 9. 2024 22:05