Development of new immunoassay platforms for rapid serological diagnosis of Lyme borreliosis

The presented work describes the development of new ligand-binding assay platforms for rapid serological diagnosis. 3-D polyethylene sinter bodies and monodisperse gold nanoparticles have been successfully employed as protein carriers, providing sensitive, selective and rapid serological diagnosis. Detection of anti-Borrelia antibodies in human sera samples was taken as a model in our work. Antibodies recognition either by visible observation or by spectroscopic measurements was easily practicable. All developed assays could be performed in a short time, 5-30 min, providing a suitable colorimetric point of care test. The established assays offer simple, rapid and reliable tools of analysis with minimal cost, which can be easily transferred to other infectious diseases

Development of new immunoassay platforms for rapid serological diagnosis of Lyme borreliosis

The presented work describes the development of new ligand-binding assay platforms for rapid serological diagnosis. 3-D polyethylene sinter bodies and monodisperse gold nanoparticles have been successfully employed as protein carriers, providing sensitive, selective and rapid serological diagnosis. Detection of anti-Borrelia antibodies in human sera samples was taken as a model in our work. Antibodies recognition either by visible observation or by spectroscopic measurements was easily practicable. All developed assays could be performed in a short time, 5-30 min, providing a suitable colorimetric point of care test. The established assays offer simple, rapid and reliable tools of analysis with minimal cost, which can be easily transferred to other infectious diseases

Surface plasmon resonance for real-time study of lectin-carbohydrate interactions for the differentiation and identification of glycoproteins.

A study of specific interactions between lectins and glycoproteins has been carried out using surface plasmon resonance (SPR) in a flow-injection mode. Lectins were covalently immobilised on the surfaces of the microfluidic sensor chip via amine coupling and serum glycoproteins were injected into the flow channels. Specific lectin-glycoprotein interactions caused the shift of refractive index proportional to the mass concentration accumulated on the channel surface. Lectins showed different affinity to the tested glycoproteins and each glycoprotein displayed its own lectin-binding pattern. It is possible to distinguish and identify even glycoproteins with similar sugar structures by simple and quick screening. The working conditions of the assay were optimised. The lectin-based SPR made it possible to carry out the label-free detection of glycoproteins within a broad concentration range with a good linearity. Regeneration conditions for the surface of the sensor chip were found and optimised. Combination of 10mM HCl and 10mM glycine-HCl (pH 2.5) removes the bound glycoproteins from the lectin surface without damaging it. The kinetic and affinity parameters of lectin-glycoprotein binding were evaluated. The proposed method was tested on human glycosylated serum. Combination of the lectin panel with SPR is suitable both for specific screening and for sensitive assay of serum glycoproteins

An integrated multiomic approach as an excellent tool for the diagnosis of metabolic diseases: our first 3720 patients

To present our experience using a multiomic approach, which integrates genetic and biochemical testing as a first-line diagnostic tool for patients with inherited metabolic disorders (IMDs). A cohort of 3720 patients from 62 countries was tested using a panel including 206 genes with single nucleotide and copy number variant (SNV/CNV) detection, followed by semi-automatic variant filtering and reflex biochemical testing (25 assays). In 1389 patients (37%), a genetic diagnosis was achieved. Within this cohort, the highest diagnostic yield was obtained for patients from Asia (57.5%, mainly from Pakistan). Overall, 701 pathogenic/likely pathogenic unique SNVs and 40 CNVs were identified. In 620 patients, the result of the biochemical tests guided variant classification and reporting. Top five diagnosed diseases were: Gaucher disease, Niemann-Pick disease type A/B, phenylketonuria, mucopolysaccharidosis type I, and Wilson disease. We show that integrated genetic and biochemical testing facilitated the decision on clinical relevance of the variants and led to a high diagnostic yield (37%), which is comparable to exome/genome sequencing. More importantly, up to 43% of these patients ( n  = 610) could benefit from medical treatments (e.g., enzyme replacement therapy). This multiomic approach constitutes a unique and highly effective tool for the genetic diagnosis of IMDs