Expanding the scope of application of clinical biochemical reagent kits for determining metabolite concentrations and enzyme activities and optimizing protocols of their using

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Introduction. The main consumers of commercial reagent kits for the study of metabolites and enzymes are clinical laboratories. Therefore, protocols for their using and calculating indices were developed mainly for blood and urine analysis. However, it is regularly necessary to determine metabolites concentration and enzymes activity to solve other tasks.

Purpose of the work – as a case study estimation of glucose concentration and amylase activity in hemolymph plasma of the gastropod mollusc Achatina fulica to demonstrate how protocols for using some clinical reagent kits for determining metabolites concentration and enzymes activity can be optimized and how the scope of their application can be expanded.

Material and Methods. The hemolymph plasma of the gastropod mollusc A. fulica served as the object of the study. Glucose concentration and amylase activity were determined in hemolymph plasma using clinical biochemical reagent kits from Spinreact (Spain). The standard and optimized protocols for determining glucose concentration are described. An optimized protocol for determining amylase activity has been described previously.

Results. A. fulica is often used as a convenient model in experimental and practical developments of different areas of focus. The knowledge of diagnostically significant biochemical indices of hemolymph plasma of mollusks makes it possible to monitor the "state of health" of animals during their cultivation in laboratories and practical using. In contrast to the original protocol for determining glucose concentration, full calibration experiments were performed, with sample analysis time reduced to one minute. The optimized protocols for using reagent kits proposed in the article were tested to determine glucose concentration and amylase activity in the hemolymph plasma of A. fulica (n=16). The values were 0,42 ± 0,07 mM and 27,8 ± 0,6 IU/L, respectively.

Conclusion. As a case study estimation of glucose concentration and amylase activity in hemolymph plasma of the gastropod mollusc Achatina fulica the effectiveness of the proposed ways of optimization of protocols for using some clinical reagent kits was demonstrated. The optimized protocols are suitable for determining the concentration of such metabolites as glucose and lactate as well as for analyzing the kinetics of various enzymes with direct measurements of their activity. The performed optimization allowed to expand the scope of application of biochemical reagent kits used in medicine, as well as to significantly reduce the time of analysis.

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作者简介

N. Malygina

St. Petersburg State University; S.M. Kirov Military Medical Academy

编辑信件的主要联系方式.
Email: n.m.malygina@mail.ru
ORCID iD: 0000-0001-9049-3969

Post-graduate Student, Department of Biochemistry, Faculty of Biology, teacher of the Department of Clinical Biochemistry and Laboratory Diagnostics

俄罗斯联邦, 7-9 Universitetskaya Emb., St Petersburg, 199034; 6, Akademika Lebedeva ul., St. Petersburg, 194044

A. Lianguzov

St. Petersburg State University

Email: andrey.lyanguzov@spbu.ru
ORCID iD: 0000-0003-4803-9070

Ph.D. (Biol.), Senior Specialist of the Observatory of Environmental Safety Resource Center of the St. Petersburg State University Science Park

俄罗斯联邦, 7-9 Universitetskaya Emb., St Petersburg, 199034

T. Petrova

St. Petersburg State University

Email: t.a.petrova@spbu.ru
ORCID iD: 0000-0002-0213-5457

Ph.D. (Biol.), Associate Professor, Specialist of the Observatory of Environmental Safety Resource Center of the St. Petersburg State University Science Park

 

俄罗斯联邦, 7-9 Universitetskaya Emb., St Petersburg, 199034

A. Ivanov

St. Petersburg State University; S.M. Kirov Military Medical Academy

Email: iamvma@mail.ru
ORCID iD: 0000-0002-8899-7524

Dr.Sc. (Med.), Professor, Corresponding Member of the RAS, Head of Department of Laboratory Medicine, Medical Institute; Head of the Department of Clinical Biochemistry and Laboratory Diagnostics President of the Association of Laboratory Specialists and Organizations «Federation of Laboratory Medicine» of Russian Federation

俄罗斯联邦, 7-9 Universitetskaya Emb., St Petersburg, 199034; 6, Akademika Lebedeva ul., St. Petersburg, 194044

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2. Figure. Calibration plot for the determining glucose concentration by the glucose oxidase method by the absorbance level of incubation medium in one minute

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