Quality control system for sample preparation and analysis results in the diagnosis of viral hepatitis B and C by real-time PCR
Abstract
Introduction. When conducting diagnostic tests for virus Infection, it is crucial to prevent the false negative results. For this, it is necessary to con-trol 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 sys-tem 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 in-to 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 In-tifica" 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, in-creases sensitivity and eliminates false…
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