Quality control system for sample preparation and analysis results in the diagnosis of viral hepatitis B and C by real-time PCR
- 作者: Moroz A.S.1,2, Toropov V.A.2, Bolshakov V.N.1,2, Aronova E.B.1
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隶属关系:
- Peter the Great St. Petersburg Polytechnic University
- Alkor Bio Group of Companies
- 期: 卷 28, 编号 3 (2025)
- 页面: 38-44
- 栏目: Biological chemistry
- URL: https://journals.eco-vector.com/1560-9596/article/view/677704
- DOI: https://doi.org/10.29296/25877313-2025-03-05
- ID: 677704
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详细
Introduction. When conducting diagnostic tests for virus Infection, it is crucial to prevent the false negative results. For this, it is necessary to control all stages of the sample preparation process and the quality of the diagnostic reaction itself.
The aim of the work is to create an optimal combination of sample preparation exogenous quality control hepatitis B and C viruses detection system using real–time multiplex PCR to improve the accuracy, reproducibility and reliability of the results.
Material and methods. Hepatitis B virus DNA and hepatitis C virus RNA isolated from blood plasma samples from patients with confirmed diagnoses were used as the object of the study. The following resources were used to perform the in silico study: the online resource Primer-BLAST, integrated into the NCBI database; the online service OligoAnalyzer Tool (Integrated DNA Technologies), as well as the programs Oligo Primer Analysis Software, Clustal Omega and UGENE. The CFX96 Touch amplifier (Bio-Rad Laboratories, USA) was used for real-time PCR. PCR was performed using the "HBV Intifica" and "HCV Intifica" kits (Alkor Bio Group of Companies, Russia).
Results. A stable and robust multiplex PCR system has been developed to control the quality of sample preparation and diagnosis of viral hepatitis B and C from human blood plasma. Application of thermodynamic analysis to primers and probes design improves the efficiency of PCR systems, increases sensitivity and eliminates false negative results.
Conclusions. Thorough optimization of viral primers and probes characteristics, together with application of exogenous internal control in multiplex reaction, significantly improves the accuracy, reproducibility and reliability of analysis results.
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作者简介
A. Moroz
Peter the Great St. Petersburg Polytechnic University; Alkor Bio Group of Companies
编辑信件的主要联系方式.
Email: anny-nice@mail.ru
ORCID iD: 0000-0002-4693-623X
SPIN 代码: 4721-1020
Post-graduate Student, Higher School of Biotechnology and Food Production; Research Engineer
俄罗斯联邦, Novorossyskaya str. 48, Saint Petersburg, 194021; Zheleznodorozhny ave., 40a, Saint Petersburg, 192148V. Toropov
Alkor Bio Group of Companies
Email: anny-nice@mail.ru
ORCID iD: 0009-0005-1993-5572
Leading Research Engineer
俄罗斯联邦, Zheleznodorozhny ave., 40a, Saint Petersburg, 192148V. Bolshakov
Peter the Great St. Petersburg Polytechnic University; Alkor Bio Group of Companies
Email: anny-nice@mail.ru
ORCID iD: 0009-0007-5126-6035
Ph.D. (Biol.), Associate Professor, Associate Professor of the Higher School of Biotechnology and Food Production; Head of the Laboratory of Molecular Diagnostics
俄罗斯联邦, Novorossyskaya str. 48, Saint Petersburg, 194021; Zheleznodorozhny ave., 40a, Saint Petersburg, 192148E. Aronova
Peter the Great St. Petersburg Polytechnic University
Email: anny-nice@mail.ru
ORCID iD: 0000-0003-4376-2972
SPIN 代码: 7288-6820
Ph.D. (Tech.), Associate Professor, Associate Professor of the Higher School of Biotechnology and Food Production
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