We use cookies to understand how you use our site and to improve your experience. This includes personalizing content and advertising. To learn more, click here. By continuing to use our site, you accept our use of cookies. Cookie Policy.

Features Partner Sites Information LinkXpress
Sign In
Advertise with Us
LGC Clinical Diagnostics

SEBIA

Sebia provides clinical protein electrophoresis equipment and reagents for in-vitro diagnostic testing, including sys... read more Featured Products: More products

Download Mobile App




Events

09 Apr 2024 - 12 Apr 2024
15 Apr 2024 - 17 Apr 2024
23 Apr 2024 - 26 Apr 2024

Semi-Automatic Isoelectric Focusing Evaluated for Apolipoprotein E Phenotyping

By LabMedica International staff writers
Posted on 30 Jan 2020
Print article
Image: The Sebia Hydragel 18 Apo E Isofocusing showing different apoE isoforms from 16 patients tested. The “control serum” provided in the Sebia kit is included in each series, together with two already typed serum samples as internal controls (as an example, E3/E2 and E4/E4) (Photo courtesy of Sorbonne University).
Image: The Sebia Hydragel 18 Apo E Isofocusing showing different apoE isoforms from 16 patients tested. The “control serum” provided in the Sebia kit is included in each series, together with two already typed serum samples as internal controls (as an example, E3/E2 and E4/E4) (Photo courtesy of Sorbonne University).
Plasma apolipoprotein E (apoE) mainly arises from liver hepatocytes (75%), but the brain (astrocytes, oligodendrocytes, microglia, astrocytes) also synthesizes apoE found in the cerebrospinal fluid.

ApoE is a major component of lipoproteins which participates in the transport and clearance of lipids. ApoE4 status is a risk factor for Alzheimer’s and other neurodegenerative diseases, whereas apoE2 and also apoE4 increase the risk for cardiovascular disease.

A team of biochemists from the Sorbonne University (Paris, France) collected blood samples from 40 patients in the course of familial hypercholesterolemia screening or mixed dyslipidemia diagnosis. These patients had given their informed consent including genetic determination and APOE genotypes were already determined by Sanger sequencing of a polymerase chain reaction (PCR) fragment encompassing the two polymorphic sites (rs429358 and rs7412).

The scientists used the Hydragel 18 Apo E Isofocusing kit (Sebia, Lisses, France) which is a qualitative kit for detection and identification of the different apoE phenotypes. A ready-to-use agarose gel containing ampholytes (pH gradient: 5–8) is used to perform a semi-automatic electrophoresis on a Sebia Hydrasys 2 Scan, followed by a specific immunofixation with anti-apoE antiserum. The “apoE isofocusing visualization” kit included antiserum diluent, stock solutions of anti-apoE antiserum and peroxidase-labeled antibody, and reagent for revelation (TTF1 and TTF2 developing solutions).

The team reported that the sera from the genotyped patients included in the study allowed them to test the following isoforms: E3/E3 (18 sera), E3/E4 (16 sera), E2/E2 (four sera) and E4/E4 (two sera). All phenotypes tested were 100% concordant to the genetic isoforms, even if all the possible phenotypes are not found in their population. Precision of the technique was verified with the repeatability and the reproducibility tests for the different isoforms, since tests gave similar results. In addition, the reproducibility tests led them to perform three freezing-thawing cycles on serum samples and a 100% concordant interpretation with genotyping was maintained. There was no interference of hemolysis, glucose or hypertriglyceridemia at the respective concentrations of 45.3 μmol/L hemoglobin, 25.1 mmol/L glucose and 9.5 and 4.8 mmol/L triglycerides for the determination of the phenotype profile.

The authors concluded that this qualitative, semi-automatized method could yield a novel and simple tool to phenotype apoE isoforms. Validated on a cohort of patient samples with known genotype, it could be used either for screening and stratification of a patient cohort, or for clinical analysis when genetic material cannot be obtained. It could be a useful and supplementary tool helping the clinician in the diagnosis of dyslipidemic and neurodegenerative diseases. The study was published in the January 2020 issue of the journal Practical Laboratory Medicine.

Related Links:
Sorbonne University
Sebia


Platinum Member
COVID-19 Rapid Test
OSOM COVID-19 Antigen Rapid Test
Specimen Collection & Transport
Anti-Cyclic Citrullinated Peptide Test
GPP-100 Anti-CCP Kit
New
Gold Member
Fully Automated Cell Density/Viability Analyzer
BioProfile FAST CDV

Print article

Channels

Molecular Diagnostics

view channel
Image: The cobas Malaria test is the first FDA-approved molecular test to screen U.S. blood donors for malaria (Photo courtesy of Roche)

First FDA-Approved Molecular Test to Screen Blood Donors for Malaria Could Improve Patient Safety

Malaria, a serious illness that often leads to death, is spread by a specific mosquito species that infect humans with a parasite. Other transmission modes include blood transfusions, organ transplants,... Read more

Hematology

view channel
Image: The low-cost portable device rapidly identifies chemotherapy patients at risk of sepsis (Photo courtesy of 52North Health)

POC Finger-Prick Blood Test Determines Risk of Neutropenic Sepsis in Patients Undergoing Chemotherapy

Neutropenia, a decrease in neutrophils (a type of white blood cell crucial for fighting infections), is a frequent side effect of certain cancer treatments. This condition elevates the risk of infections,... Read more

Pathology

view channel
Image: The medical office procedure detects the key biomarker in Parkinson’s and related neurodegenerative diseases (Photo courtesy of BIDMC)

Simple Skin Biopsy Test Detects Parkinson’s and Related Neurodegenerative Diseases

Parkinson's disease and a group of related neurodegenerative disorders known as synucleinopathies impact millions globally. These conditions, including Parkinson’s disease (PD), dementia with Lewy bodies... Read more
Copyright © 2000-2024 Globetech Media. All rights reserved.