biopreparatБИОпрепараты. Профилактика, диагностика, лечениеBiological Products. Prevention, Diagnosis, Treatment2221-996X2619-1156Scientific Centre for Expert Evaluation of Medicinal Products10.30895/2221-996X-2018-18-4-208-215biopreparat-192Research ArticleОБЗОРЫREVIEWSОпределение вирусных нуклеиновых кислот в крови человекаDetermination of Viral Nucleic Acid in the Human Bloodhttps://orcid.org/0000-0002-9402-5254АбдурашитовМ. А.AbdurashitovM. A.

р.п. Кольцово, Новосибирская область, 630559

канд. биол. наук, заведующий лабораторией эпигенетики отдела производства и применения средств ПЦР диагностики вирусных и риккетсиозных заболеваний 

Koltsovo, Novosibirsk Oblast 630559

 

Candidate of Biological Sciences, Head of the Laboratory of Epigenetics, Department of Production and Use of PCR Tools for Viral and Rickettsial Diseases Diagnostics

https://orcid.org/0000-0002-3380-9573НетесоваН. А.NetesovaN. A.

р.п. Кольцово, Новосибирская область, 630559

д-р биол. наук, заведующая отделом разработки средств ПЦР диагностикивирусных и риккетсиозных заболеваний

Koltsovo, Novosibirsk Oblast 630559

Doctor of Biological Sciences, Head of the Department of Development of PCR Tools for Viral and Rickettsial Diseases Diagnostics

ninanet@vector.nsc.ruФедеральное бюджетное учреждение науки «Государственный научный центр вирусологии и биотехнологии «Вектор» Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человекаРоссияState Research Center of Virology and Biotechnology VECTORRussian Federation201818122018184208215Copyright © Абдурашитов М.А., Нетесова Н.А., 20182018Абдурашитов М.А., Нетесова Н.А.Abdurashitov M.A., Netesova N.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.biopreparations.ru/jour/article/view/192

Многие острые вирусные инфекции вызывают сходную клиническую симптоматику, поэтому установление этиологии вирусного заболевания требует применения целых комплексов серологических или ПЦРтестов, предназначенных для выявления отдельного вида патогена. Современные методы молекулярной биологии позволяют проводить раннюю диагностику вирусных заболеваний в тот период, пока серологические методы диагностики еще не эффективны. Цель работы состояла в проведении анализа методов молекулярной диагностики, позволяющих определять вирусные нуклеиновые кислоты в крови человека. В статье представлена классификация молекулярных методов диагностики вирусных частиц в клинических образцах. Подробно рассмотрены такие методы, как гибридизация in situ, реакция обратной транскрипции (ОТ-ПЦР), двухраундовая ПЦР, мультиплексная ПЦР, а также технология ДНК-микрочипов, метод массового параллельного секвенирования. Особое внимание уделено NGS-технологиям, которые стали использоваться в вирусологии практически сразу же после их появления и позволили обнаружить целый ряд новых видов вирусов человека (включая представителей анелловирусов, пикорнавирусов, полиомавирусов и др.). Обсуждаются достоинства, проблемы, связанные с применением этих методов в клинической практике, а также перспективы их усовершенствования.

Many acute viral infections cause similar clinical symptoms, therefore, establishing the etiology of a viral disease requires the use of whole complexes of serological or PCR tests designed to detect a particular type of pathogen. Modern methods of molecular biology allow early diagnosis of viral diseases at a time when serological diagnostic methods are not yet effective. The aim of the work was to analyze molecular diagnostic methods that allow the determination of viral nucleic acids in human blood. The article presents the classification of molecular methods for the diagnosis of viral particles in clinical specimens. Methods such as in situ hybridization, reverse transcription reaction (RT-PCR), nested PCR, multiplex PCR, as well as DNA microarray technology, and the method of massive parallel sequencing are considered in detail. Particular attention is paid to NGS-technologies that were used in virology almost immediately after their appearance and allowed for detection of a number of new types of human viruses (including representatives of anelloviruses, picornaviruses, polyomaviruses, etc.). The advantages and problems associated with the application of these methods in clinical practice, as well as the prospects for their improvement are discussed.

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The authors declare that there are no conflicts of interest present.