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
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F24%3A00139615" target="_blank" >RIV/00216224:14310/24:00139615 - isvavai.cz</a>
Result on the web
<a href="https://link.springer.com/article/10.1007/s00216-024-05357-y" target="_blank" >https://link.springer.com/article/10.1007/s00216-024-05357-y</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1007/s00216-024-05357-y" target="_blank" >10.1007/s00216-024-05357-y</a>
Alternative languages
Result language
angličtina
Original language name
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
Original language description
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).
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10406 - Analytical chemistry
Result continuities
Project
<a href="/en/project/GA22-27580S" target="_blank" >GA22-27580S: Laser spectroscopy for nanometal-based label assays and imaging</a><br>
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2024
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Analytical and Bioanalytical Chemistry
ISSN
1618-2642
e-ISSN
1618-2650
Volume of the periodical
416
Issue of the periodical within the volume
30
Country of publishing house
DE - GERMANY
Number of pages
10
Pages from-to
7285-7294
UT code for WoS article
001242267600001
EID of the result in the Scopus database
2-s2.0-85195402714