Eurasian Journal of Applied Biotechnology

MODERN METHODS OF DIAGNOSTICS OF LYSOSOMAL STORAGE DISEASES

2025-07-31T11:40:16+05:00

Lysosomal storage diseases, such as Gaucher, Niemann-Pick, Pompe, Krabbe, Fabry, and Mucopolysaccharidoses, are a group of more than 50 rare inherited metabolic diseases caused by dysfunction of lysosomes due to deficiency of specific enzymes. The early and accurate diagnosis of lysosomal storage diseases is important for the correct drug administration, genetic counseling, and prevention of severe complications. In Kazakhstan, the diagnosis of lysosomal storage diseases is difficult due to limited laboratory capabilities and low prevalence of these diseases among the population. This review discusses modern diagnostic methods, including tandem mass spectrometry, next-generation sequencing, and enzymatic activity tests. For forehanded diagnosis of lysosomal storage diseases, it is important to integrate molecular, genetic, and biochemical diagnostic methods, as well as develop national newborn screening programs.

Physiological and morphological changes of heat stress in «Astana» variety of wheat plants: the role of relative water content

2026-01-21T14:01:12+05:00

Wheat (Triticum aestivum L.) is one of the most widely consumed cereal crops in the world. High temperature is one of the important abiotic stresses affecting plant growth because global warming is one of the major problems in the world today. Physiological and biochemical changes occurring in cells under abiotic stress conditions slow down plant growth and development, which ultimately leads to a decrease in wheat yield. The aim of the research was study how heat stress affects wheat growth and to find out the changes in wheat plants shape and functions under high temperature. In this study wheat of the Astana variety was used and a comparison was made between the control plants grown at 25°C and the stressed plants grown at 40°C. As a result of the work morphological and physiological changes were observed. We studied the average length of wheat roots and shoots and relative water content of Astana variety of wheat plants. In stress group roots and shoots of wheat plants were reduced in size compared to control plants and the relative water content of plants grown at optimal temperature was higher compared to plants grown under heat stress.

STRUCTURAL FEATURES OF THE CHLOROPLAST GENOME OF THE RARE TULIP SPECIES TULIPA ALBERTI, INCLUDED IN THE RED BOOK OF KAZAKHSTAN

2026-03-17T09:12:08+05:00

Tulipa alberti is an important plant species used for ornamental purposes in Asia, Europe, and North Africa. This study reports the complete chloroplast genome features of this endangered species collected from Kazakhstan using Illumina sequencing technology. The results revealed that the T. alberti chloroplast genome is highly conserved. The genome size is 152,006 bp, comprising a pair of inverted repeats (IR) of 26,330 bp, a large single-copy (LSC) region of 82,169 bp, and a small single-copy (SSC) region of 17,172 bp. The overall GC content is 36.58%. A total of 131 genes were identified, including 85 protein-coding genes, 38 tRNA genes, and 8 rRNA genes. Twenty-eight genes contained introns with lengths ranging from 540 to 2,620 bp. The nucleotide diversity (π) was 0.003257. Analysis of simple sequence repeats (SSR) identified 159 SSR loci in the T. alberti genome, mainly distributed in the LSC region (63.19%), SSC region (18.39%), and IR region (14.37%). Six types of SSR motifs (mono-, di-, tri-, tetra-, penta-, and hexanucleotides) were detected. The most polymorphic protein-coding (CDS) genes were rpoC2, cemA, rbcL, rpl36, psbH, rps3, rpl22, ndhF, ycf1, and matK, which exhibited high sequence variability (SV = 2.581–6.102) and nucleotide diversity (π = 0.004–0.010). Relative synonymous codon usage (RSCU) analysis revealed both preferred and less frequently used codons. The characterization of the T. alberti chloroplast genome provides valuable insights into the conservation of genetic resources, evolutionary and phylogenetic relationships of this rare species. However, to date, data on the complete sequencing and comprehensive bioinformatics analysis of the chloroplast genome of T. alberti remain limited. This study contributes to biodiversity preservation in Kazakhstan and offers a basis for developing effective conservation strategies at the molecular level.

OPTIMIZATION OF CULTIVATION CONDITIONS FOR RECOMBINANT INFLUENZA A/H5N1 STRAINS EXPRESSING BRUCELLA SPP. ANTIGENS (L7/L12, OMP16, OMP19, CU-ZN-SOD)

2026-03-26T10:10:04+05:00

The study presents the results of optimizing cultivation conditions for recombinant influenza A/H5N1 strains (NS1-truncated) expressing Brucella spp. antigens (L7/L12, OMP16, OMP19, CU-ZN-SOD). MDCK and Vero cell lines were used in the experiments. The effects of the infectious dose, incubation temperature, cultivation duration, as well as the effect of protease on viral replication, were determined.

It was found that the optimal conditions for cultivating recombinant influenza A strains expressing Brucella antigens are the use of MDCK cell culture, an infectious dose of 0.01 TCID50/cell, an incubation temperature of 34.0±0.5 °C, a cultivation period of 48 hours, and addition of Trypsin-TPCK at a concentration of 1.5–2.0 μg/mL. Under these conditions, viral infectivity reached ≥5.75 log TCID50/mL, and the hemagglutination titer was 1:32. The genetic stability of the viral constructs was maintained for at least four consecutive passages.

The obtained results can be used in the development and scale-up of influenza vector vaccine production technology.

OPTIMIZATION OF THE CULTURE METHOD FOR THE MOST EFFECTIVE SYNTHESIS OF POLYHYDROXYBUTYRATE BY THE BACILLUS ARYABHATTAI RAF 5 STRAIN AND ANALYSIS OF BARRIER PROPERTIES OF THE OBTAINED MEMBRANE

2025-09-30T08:52:04+05:00

The method of periodic updating (PU) or the periodic addition (PA) of fresh medium is considered as a promising technique for long-term and effective production of polyhydroxybutyrate (PHB) synthesized by different soil bacteria. In our previous studies, we reported PHB-producing potential of the Bacillus aryabhattai strain RAF 5 IMD B - 462 isolated from the chestnut soils in Astana city, Kazakhstan. The current study reports on the optimization of the RAF 5 cultivation using PU technology to obtain a high yield of PHB and biomass, as well as on the study of a membrane obtained on the basis of PHB. The accumulation of dense PHB granules in cell cytosol was confirmed by Transmission Electron Microscopy (TEM) analyses; more than 10 granules per cell were found in the cytosol of RAF 5. PHB was further extracted using the solvent extraction method. The highest PHB yield and biomass obtained after 72-hour incubation were 18.03 g/L and 20.04 g/L, respectively, when PU was applied. The highest PHB titer after 120-hour incubation was 3.76 g/L, when RAF 5 was growing in a conventional culture medium NB. The obtained PHB was further utilized to make a membrane sample which was subjected to water vapor, oxygen and carbon dioxide gas permeability analyses showing the promising results. In conclusion, this data proposes potential application might be utilized as a promising material to produce membranes.

DEVELOPMENT AND VALIDATION OF A METHOD FOR DETECTING THE BRAF V600E SOMATIC MUTATION BY THE ALLELE-SPECIFIC LAMP METHOD

2026-05-06T10:04:23+05:00

Detection of the somatic BRAF V600E mutation is a mandatory component of molecular stratification in melanoma, as its presence determines eligibility for BRAF and MEK inhibitor therapy [1–4]. Although qPCR- and NGS-based platforms remain the most widely used approaches for BRAF V600 mutation testing, interest in simpler, faster, and more cost-effective isothermal amplification formats persists [7, 22].

To develop and validate an allele-specific loop-mediated isothermal amplification (LAMP) assay for detection of the somatic BRAF V600E mutation in DNA extracted from tumour tissue.

An allele-specific LAMP format was employed. Analytical performance was characterised using linearised plasmid controls pV600WT and pV600E and their mixtures. Diagnostic validation was performed on 67 DNA samples extracted from FFPE melanoma blocks. The cobas 4800 BRAF V600 Mutation Test was used as the reference method.

The analytical limit of detection for mutant allele fraction was 1%, and the LoD₅₉ for mutant allele copy number was 200 copies per reaction. In the clinical sample set, sensitivity of LAMP-V600E relative to AS-qPCR was 93.5% (29/31), specificity 100% (36/36), positive predictive value 100%, negative predictive value 94.7%, overall accuracy 97.0%, and Cohen’s kappa coefficient 0.940. The two false-negative results were most likely attributable to FFPE-DNA degradation and the greater sensitivity of LAMP to template integrity over the comparatively longer amplification target [23, 24].

The developed allele-specific LAMP assay for BRAF V600E detection demonstrates high specificity, acceptable clinical sensitivity, and strong concordance with the reference AS-qPCR platform. The assay may be considered a rapid and technically accessible option for tissue-based BRAF testing; however, the quality and quantity of FFPE-derived DNA must be taken into account for reliable performance.