SELECTIVE BIOCHEMICAL SCREENING LYSOSOMAL STORAGE DISEASES BY TLC OF OLIGOSACCHARIDES

Mytsyk N. Y., Olkhovych N. V., Gorovenko N. G.

MEDICAL GENETICS

Scentific article

Lysosomal storage disorders (LSD) constitute a large class of hereditary metabolic diseases, including about 60 clinical entities. Aim. The excretion analysis of urine oligosaccharides using thin-layer chromatography (TLC) in patients with lysosomal storage disorders with the purpose of defining the application criteria of this method for selective biochemical screening of patients with LSD. Materials and methods. The 24-hour urine of patients (1,867 patients) with assumed hereditary metabolic diseases from different regions of Ukraine, obtained during 2004–2014, was used as the material for the study. The TLC of urine oligosaccharides was applied. Results and discussion. Oligosacchariduria, caused by different forms of LSD, is characterized by the presence of specific fractions of oligosaccharides in urine, which are not observed for healthy individuals of the respective age category. The analysis of the oligosaccharide spectrum, defined for the patients with confirmed diagnosis of LSD, was made in comparison against the control urine samples of patients, up to and exceeding 1 year of age, to exclude physiological oligosacchariduria of children of different age. The result obtained demonstrated that the spectrum of fractions of oligosaccharides in urine in the samples of patients with LSD and their Rlac value was considerably different from the samples of healthy subjects of different age categories, which allowed using the TLC method for urine oligosaccharides with the purpose of selective biochemical screening of patients with assumed lysosomal pathology. The patients with sialidosis had specific fractions of oligosaccharides in the region of Rlac 0.20 and 0.75 with no other fractions present. In case of α-mannosidosis there was evident excretion of a number of oligosaccharides, different in their mobility, with the Rlac value from 0.043 to 0.7, which is not remarkable for healthy individuals of the older age category. The TLC plates of patients with GM1-gangliosidosis, galactosialidosis and MPS І, ІІ and IV B type had remarkable evident and well-stained groups of pathological fractions of oligosaccharides with the value of Rlac<0.15, which correspond to hepta- and octasaccharides, not present in healthy individuals of all the age groups. However, the spectra of oligosaccharides, revealed in these patients, were similar among themselves, which did not allow distinguishing these diseases. The pathological fractions of hepta- and octasaccharides are common for several lysosomal diseases, thus, while revealing the latter it is possible only to assume the presence of LSD in a patient. Therefore, it is obligatory for all the cases of revealing the increased excretion of pathological oligosaccharides with urine to conduct confirmatory diagnostics with the determination of the primary biochemical deficiency, which resulted in the accumulation of these metabolites. Thus, thin-layer chromatography of urine oligosaccharides is available for wide application in specialized laboratories using the method, which allows using it in the course of selective biochemical screening of patients with LSD, accompanied with oligosacchariduria, and, as a result, diminishing the probability of false diagnostics of hereditary diseases and providing for more reasonable application of expensive methods of confirmatory diagnostics. Conclusions. The TLC method of urine oligosaccharides allows narrowing the search groups of patients who are assumed to have LSD, thus it may be recommended for application in the course of selective biochemical screening of patients of LSD, accompanied with oligosacchariduria.

oligosaccharides, TLC, oligosacchariduria, lysosomal diseases

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«Bulletin of problems biology and medicine» Issue 1 part 1 (126), 2016 year, 222-227 pages, index UDK 616-056.7-07