Influence of preanalytical stage violations on the reliability of the result of SARS-CoV-2 diagnostics by real-time polymerase chain reaction method
- Authors: Gusyakova O.A.1, Antipov V.A.1, Avakyan N.A.2, Medvedeva A.S.1, Kartashov V.V.1
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Affiliations:
- Samara State Medical University
- State Budgetary Health Care Institution of the Moscow Region "Center for the Prevention and Control of AIDS"
- Issue: Vol 28, No 10 (2025)
- Pages: 21-26
- Section: Biological chemistry
- URL: https://journal-vniispk.ru/1560-9596/article/view/334351
- DOI: https://doi.org/10.29296/25877313-2025-10-03
- ID: 334351
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Abstract
Introduction. COVID-19 laboratory testing by polymerase chain reaction is a key step for making the correct diagnosis and prescribing the most effective treatment. Each laboratory testing includes a preanalytical phase, compliance with which is necessary to fulfill the global task of obtaining a reliable result. According to the interim guidelines for laboratory diagnostics of COVID-19 in a pandemic, swabs from the nasopharynx or oropharynx in a sterile plastic tube with a transport medium are transported at a temperature of +2 to +8 °C. The time of transportation of samples to the laboratory should not exceed 5 days, and long-term storage is allowed at –20 °C or –70 °C.
The aim of the study is to evaluate the effect of physical factors that do not meet the preanalytical phase requirements on reliability of SARS-CoV-2 RNA detection by real-time polymerase chain reaction.
Material and Methods. Fifty SARS-CoV-2 positive respiratory swabs were utilized, each of which was divided into 4 samples (groups): first group of samples was incubated for 7 days at an acceptable temperature (+5 °C), the second group – 7 days at an unacceptable temperature (+25 °C), third group was subjected to double freezing, and fourth group was exposed to ultraviolet radiation. Then, the samples were re-tested by polymerase chain reaction in real time and the threshold amplification cycle was interpreted.
Results. The influence of ultraviolet radiation led to a decrease in the number of positive samples by 20%, as well as to an increase in the median of the threshold amplification cycle by 1.1 units. Upon double freezing of biological material, a 18% decrease in the proportion of positive samples was observed. Prolonged storage of samples at +5 °C led to a 20% decrease, while storage at +25 °C resulted in a 14% decrease. There were no significant differences between the threshold cycle of experimental and control samples for the given factors.
Conclusions. Among the examined physical factors, ultraviolet radiation has the most significant negative impact on the preservation of SARS-CoV-2 RNA. Prolonged storage of samples at +5 ° C and +25 °C, similarly to double freezing of clinical material, also has a negative effect.
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##article.viewOnOriginalSite##About the authors
O. A. Gusyakova
Samara State Medical University
Author for correspondence.
Email: o.a.gusyakova@samsmu.ru
ORCID iD: 0000-0002-5619-4583
SPIN-code: 5198-8830
Dr.Sc. (Med.), Professor, Head of the Department of Fundamental and Clinical Biochemistry with Laboratory Diagnostics
Russian Federation, 89 Chapaevskaya str., Samara, 443099V. A. Antipov
Samara State Medical University
Email: v.a.antipov@samsmu.ru
ORCID iD: 0000-0002-5295-3554
SPIN-code: 9507-1324
Assistant of the Department of Fundamental and Clinical Biochemistry with Laboratory Diagnostics, Biologist of the Laboratory of Development and Expertise of New Medical Devices for in vitro Diagnostics of the Professional Center for Education and Research in Genetic and Laboratory Technologies
Russian Federation, 89 Chapaevskaya str., Samara, 443099N. A. Avakyan
State Budgetary Health Care Institution of the Moscow Region "Center for the Prevention and Control of AIDS"
Email: lol_0310@mail.ru
ORCID iD: 0000-0003-1174-223X
Doctor of Clinical Laboratory Diagnostics
Russian Federation, 41а Silikat microdistrict, Kotelniki, Moscow region, 140053A. S. Medvedeva
Samara State Medical University
Email: an.s.medvedeva@samsmu.ru
ORCID iD: 0009-0005-1041-9678
SPIN-code: 8999-4275
Assistant of the Department of Fundamental and Clinical Biochemistry with Laboratory Diagnostics
Russian Federation, 89 Chapaevskaya str., Samara, 443099V. V. Kartashov
Samara State Medical University
Email: stomkvv@gmail.com
ORCID iD: 0000-0002-8671-2898
SPIN-code: 7554-8410
Assistant of the Department of Orthopedic Dentistry
Russian Federation, 89 Chapaevskaya str., Samara, 443099References
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