The pros and cons of nucleic acid-amplified immunoassays—a
comparative study on the quantitation of prostate-specific
antigen with and without rolling circle amplification

J 2024

The pros and cons of nucleic acid-amplified immunoassays—a comparative study on the quantitation of prostate-specific antigen with and without rolling circle amplification

DEKALIUK, Mariia; Zdeněk FARKA a Niko HILDEBRANDT

Základní údaje

Originální název

The pros and cons of nucleic acid-amplified immunoassays—a comparative study on the quantitation of prostate-specific antigen with and without rolling circle amplification

Autoři

DEKALIUK, Mariia; Zdeněk FARKA a Niko HILDEBRANDT

Vydání

Analytical and Bioanalytical Chemistry, Heidelberg, Springer Nature, 2024, 1618-2642

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10406 Analytical chemistry

Stát vydavatele

Německo

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 3.800

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14310/24:00139615

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

ELISA; Diagnostics; PSA; Fluorescence; Terbium; TR-FRET

Štítky

14313050, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 5. 2. 2025 11:50, Mgr. Marie Novosadová Šípková, DiS.

Anotace

V originále

Integrating isothermal nucleic acid amplification strategies into immunoassays can significantly decrease analytical limits of detection (LODs). On the other hand, an amplification step adds time, complication, reagents, and costs to the assay format. To evaluate the pros and cons in the context of heterogeneous multistep immunoassays, we quantified prostate-specific antigen (PSA) with and without rolling circle amplification (RCA). In addition, we compared time-gated (TG) with continuous-wave (CW) photoluminescence (PL) detection using a terbium complex and a fluorescein dye, respectively. For both direct (non-amplified) and amplified assays, TG PL detection provided circa four- to eightfold lower LODs, illustrating the importance of autofluorescence background suppression even for multi-wash assay formats. Amplified assays required an approximately 2.4 h longer assay time but led to almost 100-fold lower LODs down to 1.3 pg/mL of PSA. Implementation of TG-FRET (using a Tb-Cy5.5 donor–acceptor pair) into the RCA immunoassay resulted in a slightly higher LOD (3.0 pg/mL), but the ratiometric detection format provided important benefits, such as higher reproducibility, lower standard deviations, and multiplexing capability. Overall, our direct comparison demonstrated the importance of biological background suppression even in heterogeneous assays and the potential of using isothermal RCA for strongly decreasing analytical LODs, making such assays viable alternatives to conventional enzyme-linked immunosorbent assays (ELISAs).

Návaznosti

GA22-27580S, projekt VaV

Název: Laserová spektroskopie v imunostanovení a zobrazování s nanometalickými značkami (Akronym: IMMUNOLIBS)

Investor: Grantová agentura ČR, Laserová spektroskopie v imunostanovení a zobrazování s nanometalickými značkami

Citovat

DEKALIUK, Mariia; Zdeněk FARKA a Niko HILDEBRANDT. The pros and cons of nucleic acid-amplified immunoassays—a comparative study on the quantitation of prostate-specific antigen with and without rolling circle amplification. Analytical and Bioanalytical Chemistry. Heidelberg: Springer Nature, 2024, roč. 416, č. 30, s. 7285-7294. ISSN 1618-2642. Dostupné z: https://doi.org/10.1007/s00216-024-05357-y.

@article{2455123,
   author = {Dekaliuk, Mariia and Farka, Zdeněk and Hildebrandt, Niko},
   article_location = {Heidelberg},
   article_number = {30},
   doi = {https://doi.org/10.1007/s00216-024-05357-y},
   keywords = {ELISA; Diagnostics; PSA; Fluorescence; Terbium; TR-FRET},
   language = {eng},
   issn = {1618-2642},
   journal = {Analytical and Bioanalytical Chemistry},
   title = {The pros and cons of nucleic acid-amplified immunoassays—a comparative study on the quantitation of prostate-specific antigen with and without rolling circle amplification},
   url = {https://link.springer.com/article/10.1007/s00216-024-05357-y},
   volume = {416},
   year = {2024}
}

TY  - JOUR
ID  - 2455123
AU  - Dekaliuk, Mariia - Farka, Zdeněk - Hildebrandt, Niko
PY  - 2024
TI  - The pros and cons of nucleic acid-amplified immunoassays—a comparative study on the quantitation of prostate-specific antigen with and without rolling circle amplification
JF  - Analytical and Bioanalytical Chemistry
VL  - 416
IS  - 30
SP  - 7285-7294
EP  - 7285-7294
PB  - Springer Nature
SN  - 16182642
KW  - ELISA
KW  - Diagnostics
KW  - PSA
KW  - Fluorescence
KW  - Terbium
KW  - TR-FRET
UR  - https://link.springer.com/article/10.1007/s00216-024-05357-y
N2  - Integrating isothermal nucleic acid amplification strategies into immunoassays can significantly decrease analytical limits of detection (LODs). On the other hand, an amplification step adds time, complication, reagents, and costs to the assay format. To evaluate the pros and cons in the context of heterogeneous multistep immunoassays, we quantified prostate-specific antigen (PSA) with and without rolling circle amplification (RCA). In addition, we compared time-gated (TG) with continuous-wave (CW) photoluminescence (PL) detection using a terbium complex and a fluorescein dye, respectively. For both direct (non-amplified) and amplified assays, TG PL detection provided circa four- to eightfold lower LODs, illustrating the importance of autofluorescence background suppression even for multi-wash assay formats. Amplified assays required an approximately 2.4 h longer assay time but led to almost 100-fold lower LODs down to 1.3 pg/mL of PSA. Implementation of TG-FRET (using a Tb-Cy5.5 donor–acceptor pair) into the RCA immunoassay resulted in a slightly higher LOD (3.0 pg/mL), but the ratiometric detection format provided important benefits, such as higher reproducibility, lower standard deviations, and multiplexing capability. Overall, our direct comparison demonstrated the importance of biological background suppression even in heterogeneous assays and the potential of using isothermal RCA for strongly decreasing analytical LODs, making such assays viable alternatives to conventional enzyme-linked immunosorbent assays (ELISAs).
ER  -

DEKALIUK, Mariia; Zdeněk FARKA a Niko HILDEBRANDT. The pros and cons of nucleic acid-amplified immunoassays—a comparative study on the quantitation of prostate-specific antigen with and without rolling circle amplification. \textit{Analytical and Bioanalytical Chemistry}. Heidelberg: Springer Nature, 2024, roč.~416, č.~30, s.~7285-7294. ISSN~1618-2642. Dostupné z: https://doi.org/10.1007/s00216-024-05357-y.

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