Related Experiment Video

Updated: Jan 29, 2026

Bidirectional Retroviral Integration Site PCR Methodology and Quantitative Data Analysis Workflow
12:53

Bidirectional Retroviral Integration Site PCR Methodology and Quantitative Data Analysis Workflow

Published on: June 14, 2017

11.2K

Methodological aspects of Universal immuno-PCR on standard tubes

J E Abud1, C G Santamaría1, M Oggero2

  • 1Instituto de Salud y Ambiente Del Litoral ISAL, CONICET-UNL, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional Del Litoral, Ciudad Universitaria, Ruta Nacional N°168, km 472, CPA S3000ZAA, Santa Fe, Argentina.

Analytical Biochemistry
|February 16, 2019
PubMed

Insights

Researchers improved the human thyroid-stimulating hormone (hTSH) immuno-polymerase chain reaction assay using a pre-assembled DNA-neutravidin complex. This enhancement boosted assay sensitivity and reproducibility for more accurate results.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Assay Development

Background:

  • The "Universal" immuno-polymerase chain reaction (IPCR) format commonly uses biotinylated DNA, IgG, and avidin conjugates.
  • Optimizing IPCR assays is crucial for enhancing diagnostic sensitivity and reproducibility.

Purpose of the Study:

  • To evaluate methods for improving the analytical performance of the human thyroid-stimulating hormone (hTSH)-IPCR assay.
  • To investigate the utility of mono- and bi-biotinylated DNA probes, pre-self-assembled DNA-neutravidin complexes, blocking steps, and glutaraldehyde pretreatment.

Main Methods:

  • Utilized mono- and bi-biotinylated DNA probes in the hTSH-IPCR assay.
  • Investigated pre-self-assembled DNA-neutravidin complexes.
  • Assessed the impact of blocking steps and glutaraldehyde pretreatment on assay performance.
  • Compared the optimized hTSH-IPCR assay with self-made and commercial ELISA kits.

Main Results:

  • The pre-self-assembled mono-biotinylated DNA-neutravidin complex significantly enhanced sensitivity and reproducibility.
  • The optimized hTSH-IPCR assay achieved an improved limit of detection (LOD) of 0.01 μIU/ml.
  • Superior calibration sensitivity (SEN: 2.4) and analytic sensitivity (γ: 9 μIU/ml-1) were observed compared to ELISA methods.

Conclusions:

  • Pre-self-assembled mono-biotinylated DNA-neutravidin complexes offer a robust strategy for improving hTSH-IPCR assay performance.
  • This approach enhances assay sensitivity and reproducibility, even without a blocking step.
  • The developed hTSH-IPCR assay demonstrates superior analytical performance over traditional ELISA methods.

Keywords:
DiagnosticHuman TSHImmune-PCRImmunoassaysMonoclonal antibodies

More Related Videos

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells10:07

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

Published on: August 25, 2017

8.4K
Peptide and Protein Quantification Using Automated Immuno-MALDI iMALDI08:57

Peptide and Protein Quantification Using Automated Immuno-MALDI iMALDI

Published on: August 18, 2017

8.3K

Related Concept Videos

PCR01:32

PCR

Overview
237.3K
Measurement: Standard Units03:38

Measurement: Standard Units

Every measurement provides three kinds of information: the size or magnitude of the measurement (a number), a standard of comparison for the measurement (a unit), and an indication of the uncertainty of the measurement. While the number and unit are explicitly represented when a quantity is written, the uncertainty is an aspect of the errors in the measurement results.
79.4K
Uterine Tubes01:16

Uterine Tubes

The uterine or fallopian tubes function as the conduit through which oocytes travel from the ovaries to the uterus. Each fallopian tube measures approximately 10 to 13 cm long and is anatomically divided into the infundibulum, ampulla, isthmus, and interstitial part (or intramural segment). The infundibulum is characterized by its funnel shape and features extensions called fimbriae which reach towards the peritoneal cavity. These fimbriae play a critical role during ovulation as they extend...
2.3K
Quantitative Aspects of Drug-Receptor Interaction01:30

Quantitative Aspects of Drug-Receptor Interaction

The receptor occupancy theory connects a drug's response to the number of occupied receptors. With higher drug concentrations, more receptors are occupied, leading to increased responses. The formation of drug-receptor complexes involves association and dissociation rates, which reach equilibrium when the forward and backward reactions are equal. The equilibrium association constant (Ka) and its inverse, the equilibrium dissociation constant (Kd), indicate drug affinity. Higher Ka and lower...
1.8K
Endotracheal Tube Extubation01:24

Endotracheal Tube Extubation

Endotracheal tube extubation is a critical procedure in weaning patients from mechanical ventilation. It involves physically removing the oral or nasal endotracheal (ET) tube, marking the final step in liberating a patient from ventilatory support.
Procedure
Extubation removes the endotracheal tube (ETT) from the patient on mechanical ventilation. It requires a well-coordinated, multidisciplinary approach involving physicians, nurses, respiratory therapists, and other healthcare professionals....
4.0K
Standard Enthalpy of Formation02:37

Standard Enthalpy of Formation

Enthalpy changes are typically tabulated for reactions in which both the reactants and products are at the same conditions. A standard state is a commonly accepted set of conditions used as a reference point for the determination of properties under other different conditions. For chemists, the IUPAC standard state refers to materials under a pressure of 1 bar and solutions at 1 M and does not specify a temperature. Many thermochemical tables list values with a standard state of 1 atm. Because...
49.1K