ELISA Sensitivity and Specificity

High-Affinity Antibodies:

Finding antibodies that bind strongly improves ELISA sensitivity but can be difficult and time-consuming because of natural variations in immune responses.

Low-Affinity Antigens:

Using antigens with weaker binding allows for flexible sensitivity adjustments but can raise safety concerns for those handling them.

Signal Boosting:

Methods like using nanomaterials or enzymes increase signal strength but can also increase background noise, reducing signal-to-noise ratios.

New Signal Systems:

New ways to identify signals can improve the variety of signals detected but may require investment in new signal acquisition technologies.

More HRP in Kits:

Commercial ELISA kits increase sensitivity by adding more HRP conjugation to antibodies, allowing for more precise detection at lower levels.

Estimating Detection Limits:

Using the antibody-antigen binding constant from homogeneous solutions is important for estimating the detection limit, as demonstrated by Jia et al., who achieved a 25-fold lower limit for p53 cancer biomarkers with MMP-based assays and AuNP amplification.

Microfluidic ELISA:

Innovative microfluidic ELISA improves sensitivity via analyte preconcentration, making it easier to detect smaller amounts. However, it can be limited by antibodies getting saturated in certain areas.

Digital Immunoassays:

These use tiny beads to increase sensitivity and speed, with amplified signals detected by flow cytometers.

Recombinant B-Cell Multi-Epitopes:

Creating these multi-epitopes improves the strength and accuracy of antigens, paving the way for advanced indirect ELISA kits for diagnosing human fascioliasis.

Customized ELISA in Racing Labs:

Customizing ELISA tests in racing laboratories increases accuracy by reducing false positives and improving drug screening.

Nucleic Acid Amplification Assays:

qRT-PCR is very sensitive and accurate for detecting arbovirus infections. However, their clinical usefulness is limited by short infection periods and varying detection rates in different sample types like spinal fluid and serum.

Aptamer Specificity in ELISA:

Improving aptamer accuracy involves techniques like altering binding properties, removing non-specific variants, and selecting precise targets to ensure reliable results.

Advanced Antibody Strategies in Proteomics:

Methods like SRM (Selected Reaction Monitoring) and PEA (Proximity Extension Assay) use special antibodies to improve accuracy in detecting biomarkers, crucial for identifying disease pathways and developing treatments.