AI Article Synopsis
- A new ELISA method called DLISA was created using DNAzymes, which are more stable than traditional enzyme-based methods, allowing for better detection in harsh environments.
- This method utilizes a combination of catalytic and molecular beacons to achieve ultrasensitive detection of proteins like human IgG, with a detection limit as low as 2 fg/mL.
- DLISA is also capable of multiplex detection, meaning it can identify multiple targets at once, making it a promising low-cost and high-throughput tool for clinical diagnostics.
Article Abstract
A DNAzyme-based ELISA, termed DLISA, was developed as a novel protein enzyme-free, triply amplified platform, combining a catalytic and molecular beacon (CAMB) system with a cation exchange reaction for ultrasensitive multiplex fluorescent immunosorbent assay. Classical ELISA, which employs protein enzymes as biocatalysts to afford amplified signals, suffers from poor stability caused by the irreversible denaturation of these enzymes under harsh conditions, such as heat and acidity. Compared with proteins, nucleic acids are more stable and adaptable, and they can be easily produced using a commercial DNA synthesizer. Moreover, the catalytic and cleavage activities of DNAzyme can be achieved in solution; thus, no enzyme immobilization is needed for detection. Taken together, these attributes suggest that a DNAzyme-based ELISA detection approach will be more robust than current ELISA assays. Importantly, the proposed triply amplified DLISA immunoassay method shows ultrasensitive detection of such targets as human IgG with a detection limit of 2 fg/mL (3 × 10(-17) M), which is well within the range of many important disease biomarkers. DLISA can also be used to construct a sensing array for simultaneous multiplexed detection. With these merits, this high-throughput, stable, simple, sensitive, and low-cost multiplex fluorescence immunoassay shows promise for applications in clinical diagnosis.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1021/acs.analchem.5b01323 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
A Novel DNAzyme-Based Fluorescent Biosensor for Detection of RNA-Containing Nipah Henipavirus.
Biosensors (Basel)
February 2023
Saint-Petersburg Pasteur Institute, 197101 Saint-Petersburg, Russia.
Nipah virus (NiV) is a zoonotic RNA virus which infects humans and animals in Asian countries. Infection in humans occurs in different forms, from asymptomatic infection to fatal encephalitis, and death occurred in 40-70% of those infected in outbreaks that occurred between 1998 and 2018. Modern diagnostics is carried out by real-time PCR to identify pathogens or by ELISA to detect antibodies.
View Article and Find Full Text PDFSimultaneous Determination of Antibiotics, Mycotoxins, and Hormones in Milk by an 8-17 DNAzyme-Based Enzyme-Linked Immunosorbent Assay.
J Agric Food Chem
October 2022
State Key Laboratory of Food Science and Technology, National Centre for Technology Innovation on Fast Biological Detection of Grain Quality and Safety, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China.
The simultaneous detection of three kinds of small-molecule contaminants (antibiotics, mycotoxins, and hormones) in milk was realized by using an 8-17 DNAzyme-based fluorescent enzyme-linked immunosorbent assay (ELISA), in which 8-17 DNAzyme was utilized as the catalytic enzyme for amplifying the signal. Compared with the conventional ELISA in which horseradish peroxidase is used as the catalyzing factor, this 8-17 DNAzyme-based ELISA could achieve multicolor signal output with lower detection limits. The linearities for chloramphenicol, 17β-estradiol, and aflatoxin M were in the range of 0.
View Article and Find Full Text PDFHybridization chain reaction and DNAzyme-based dual signal amplification strategy for sensitive fluorescent sensing of aflatoxin B1 by using the pivot of triplex DNA.
Food Res Int
August 2022
School of Chemistry, Sun Yat-Sen University, Guangzhou 510275, PR China.
This work developed an enzyme-free fluorescent aptasensor for sensitive aflatoxin B1 (AFB1) detection based on a dual signal amplification strategy of hybridization chain reaction (HCR) and Zn-dependent DNAzyme. In the presence of AFB1, the aptamer specifically binds to the target, releasing the blocking DNA, which can initiate HCR between hairpin probes H1 and H2. With the addition of the substrate strand (Zn-Sub) and enzyme strand (Zn-Enz) of DNAzyme, HCR product can hybridize with Zn-Sub and Zn-Enz to form triplex DNA and Y-shaped structure together, which further activates the DNAzyme to cleave Zn-Sub.
View Article and Find Full Text PDFHoming peptide combined with DNAzyme-based ELISA-like assay for highly specific and sensitive detection of fibrin.
Talanta
February 2022
Shandong Provincial Key Laboratory of Chemical Energy Storage and Novel Cell Technology, School of Chemistry and Chemical Engineering, Liaocheng University, Liaocheng, 252000, China. Electronic address:
A highly sensitive and specific ELISA-like chemiluminescence method for detection of fibrin has been developed. In the sensing platform, the homing peptide (CREKA), as recognition molecule, which can specially recognize the fibrin on microtiter plate, combined with G-quadruplex-based DNAzyme to form the probe of G-quadruplex-hemin DNAzyme-CREKA. After the sample solution was coated on the plates, the probe was crosslinked with fibrin through the interaction of CREKA and fibrin.
View Article and Find Full Text PDFDLISA: A DNAzyme-Based ELISA for Protein Enzyme-Free Immunoassay of Multiple Analytes.
Anal Chem
August 2015
†Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Biology, Collaborative Innovation Center for Molecular Engineering for Theranostics, Hunan University, Changsha 410082, China.
Article Synopsis
- A new ELISA method called DLISA was created using DNAzymes, which are more stable than traditional enzyme-based methods, allowing for better detection in harsh environments.
- This method utilizes a combination of catalytic and molecular beacons to achieve ultrasensitive detection of proteins like human IgG, with a detection limit as low as 2 fg/mL.
- DLISA is also capable of multiplex detection, meaning it can identify multiple targets at once, making it a promising low-cost and high-throughput tool for clinical diagnostics.
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!