Nonlinear hybridization chain reaction-based flow cytometric immunoassay for the detection of prostate specific antigen

Zeng, Zhuoer; Zhou, Rong; Sun, Ruowei; Zhang, Xun; Zhang, Di; Zhu, Qubo; Chen, Chuanpin

doi:10.1016/j.aca.2022.340048

Cited by 9 publications

(6 citation statements)

References 40 publications

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“…Due to its high specificity and wide applicability, immunoassay has been popular in disease diagnosis, environmental safety monitoring, and drug efficacy evaluation. , Thereinto, the ultrasensitive immunoassay for the detection of low levels of protein markers is essential for the early diagnosis and treatment of diseases. , Currently, the sensitive immunodetection of trace proteins usually requires the help of signal amplification strategies, mainly including nucleic acid amplification technique, − enzyme-assisted amplification, − etc. However, these amplification techniques usually rely on some sophisticated instruments or equipment in specialized central laboratories and are complex, time-consuming, and not easy to popularize among the public, especially for the applications in resource-limited regions. , …”

Section: Introductionmentioning

confidence: 99%

Tyramine-Invertase Bioconjugate-Amplified Personal Glucose Meter Signaling for Ultrasensitive Immunoassay

Dai,

Zhang,

et al. 2024

Anal. Chem.

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Highly sensitive and facile detection of low levels of protein markers is of great significance for the early diagnosis and efficacy monitoring of diseases. Herein, aided by an efficient tyramine-signal amplification (TSA) mechanism, we wish to report a simple but ultrasensitive immunoassay with signal readout on a portable personal glucose meter (PGM). In this study, the bioconjugates of tyramine and invertase (Tyr-inv), which act as the critical bridge to convert and amplify the protein concentration information into glucose, are prepared following a click chemistry reaction. Then, in the presence of a target protein, the sandwich immunoreaction between the immobilized capture antibody, the target protein, and the horseradish peroxidase (HRP)-conjugated detection antibody is specifically performed in a 96-well microplate. Subsequently, the specifically loaded HRP-conjugated detection antibodies will catalyze the amplified deposition of a large number of Tyr-inv molecules onto adjacent proteins through highly efficient TSA. Then, the deposited invertase, whose dosage can faithfully reflect the original concentration of the target protein, can efficiently convert sucrose to glucose. The amount of finally produced glucose is simply quantified by the PGM, realizing the highly sensitive detection of trace protein markers such as the carcinoembryonic antigen and alpha fetoprotein antigen at the fg/mL level. This method is simple, cost-effective, and ultrasensitive without the requirement of sophisticated instruments or specialized laboratory equipment, which may provide a universal and promising technology for highly sensitive immunoassay for in vitro diagnosis of diseases.

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Section: Introductionmentioning

confidence: 99%

Tyramine-Invertase Bioconjugate-Amplified Personal Glucose Meter Signaling for Ultrasensitive Immunoassay

Dai,

Zhang,

et al. 2024

Anal. Chem.

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“…[14][15][16] The hybridization chain reaction (HCR) developed by Dirks and Pierce is an enzyme-free isothermal amplication technique that is widely used in nucleic acid and cancer cell analyses as well as protein detection applications. [17][18][19][20][21][22][23] In this approach, nickel DNA nanowires are polymerized using an initiator sequence with chain displacement between two hairpins. There are notable drawbacks to the traditional HCR process, including linear growth and slow kinetics that limit its sensitivity in clinical diagnosis; there is also persistent background leakage even in the absence of initiators due to the spontaneous opening of hairpin probes.…”

Section: Introductionmentioning

confidence: 99%

“…However, few previous researchers have attempted to use hairpin-free NHCR for protein detection. 16,23,31,32 Magnetic beads (MBs) are widely used in immunosensor construction for clinical diagnosis, environmental monitoring, and food analysis as they are advantageous components in immunosensors. 33,34 For instance, the surfaces of MBs are highly modiable thus allowing for antigen-antibody interactions.…”

Section: Introductionmentioning

confidence: 99%

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Rational design of nonlinear hybridization immunosensor chain reactions for simultaneous ultrasensitive detection of two tumor marker proteins

Zeng

Zhou

et al. 2023

Anal. Methods

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“…Therefore, accurate diagnosis and specific treatment of early-stage PCa are crucial for male health [3,4]. Prostate-specific antigen (PSA), a primary biomarker for PCa [5,6], necessitates rapid and highly sensitive detection for effective screening and early diagnosis [7], [8,9]. To date, a range of assays has been developed for the detection of PCa biomarkers, encompassing fluorescence immunoassays [10], photoelectrochemical methods [11,12], and electrochemical approaches [13,14].…”

Section: Introductionmentioning

confidence: 99%

Digital droplet immunoassay based on a microfluidic chip with magnetic beads for the detection of prostate‐specific antigen

Zeng,

Tian,

Ren

et al. 2023

J of Separation Science

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Sensitive biomarker detection techniques are beneficial for both disease diagnosis and postoperative examinations. In this study, we report an integrated microfluidic chip designed for the immunodetection of prostate‐specific antigens (PSAs). The microfluidic chip is based on the three‐dimensional structure of quartz capillaries. The outlet channel extends to 1.8 cm, effectively facilitating the generation of uniform droplets ranging in size from 3 to 50 μm. Furthermore, we successfully immobilized the captured antibodies onto the surface of magnetic beads using an activator, and we constructed an immunosandwich complex by employing biotinylated antibodies. A key feature of this microfluidic chip is its integration of microfluidic droplet technology advantages, such as high‐throughput parallelism, enzymatic signal amplification, and small droplet size. This integration results in an exceptionally sensitive PSA detection capability, with the detection limit reduced to 7.00 ± 0.62 pg/mL.

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Nonlinear hybridization chain reaction-based flow cytometric immunoassay for the detection of prostate specific antigen

Cited by 9 publications

References 40 publications

Tyramine-Invertase Bioconjugate-Amplified Personal Glucose Meter Signaling for Ultrasensitive Immunoassay

Tyramine-Invertase Bioconjugate-Amplified Personal Glucose Meter Signaling for Ultrasensitive Immunoassay

Rational design of nonlinear hybridization immunosensor chain reactions for simultaneous ultrasensitive detection of two tumor marker proteins

Digital droplet immunoassay based on a microfluidic chip with magnetic beads for the detection of prostate‐specific antigen

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