veterinaryВетеринария сегодняVeterinary Science Today2304-196X2658-6959"Veinard"10.29326/2304-196X-2023-12-3-197-207veterinary-739Research ArticleОБЗОРЫ | ОБЩИЕ ВОПРОСЫREVIEWS | GENERAL ISSUESСовременные подходы к разработке тест-систем на основе количественной ПЦР в режиме реального времениCurrent approaches to development of real-time qPCR test-kitshttps://orcid.org/0000-0002-4682-6559ДоронинМ. И.DoroninM. I.

Доронин Максим Игоревич, доктор биологических наук, заведующий сектором лаборатории профилактики ящура

г. Владимир

Maksim I. Doronin, Doctor of Science (Biology), Head of Sector, Laboratory for FMD Prevention

Vladimir

doronin@arriah.ruhttps://orcid.org/0000-0003-1718-1955МихалишинД. В.MikhalishinD. V.

Михалишин Дмитрий Валерьевич, доктор ветеринарных наук, заведующий лабораторией профилактики ящура

г. Владимир

Dmitry V. Mikhalishin, Doctor of Science (Veterinary Medicine), Head of Laboratory for FMD Prevention

Vladimir

mihalishindv@arriah.ruhttps://orcid.org/0000-0001-5982-3675СпрыгинА. В.SpryginA. V.

Спрыгин Александр Владимирович, доктор биологических наук, старший научный сотрудник референтной лаборатории болезней крупного рогатого скота

г. Владимир

Alexander V. Sprygin, Doctor of Science (Biology), SeniorResearcher, Reference Laboratory for Bovine Diseases

Vladimir

sprygin@arriah.ruhttps://orcid.org/0000-0002-5982-8393МазлумА.MazloumA.

Мазлум Али, кандидат биологических наук, старший научный сотрудник референтной лаборатории по африканской чуме свиней

г. Владимир

Ali Mazloum, Candidate of Science (Biology), Senior Researcher, Reference Laboratory for African Swine Fever

Vladimir

mazlum@arriah.ruhttps://orcid.org/0000-0002-9857-5915ЖбановаТ. В.ZhbanovaT. V.

Жбанова Татьяна Валентиновна, кандидат биологических наук, младший научный сотрудник отдела образования и научной информации, ученый секретарь диссертационногосовета

г. Владимир

Tatyana V. Zhbanova, Candidate of Science (Biology), Junior Researcher, Education and Scientific Support Department, Academic Secretary of Thesis Council

Vladimir

zhbanova@arriah.ruhttps://orcid.org/0000-0003-3159-1969ГруздевК. Н.GruzdevK. N.

Груздев Константин Николаевич, доктор биологических наук, профессор, главный научный сотрудник информационно-аналитического центра

г. Владимир

Konstantin N. Gruzdev, Doctor of Science (Biology), Professor, Chief Researcher, Information and Analysis Centre

Vladimir

gruzdev@arriah.ruhttps://orcid.org/0000-0003-4054-8532ЧернышоваЕ. В.ChernyshovaE. V.

Чернышова Елена Владимировна, кандидат ветеринарных наук, младший научный сотрудник референтной лаборатории по бешенству и BSE

г. Владимир

Elena V. Chernyshova, Candidate of Science (VeterinaryMedicine), Junior Researcher, Reference Laboratory for Rabies and BSE

Vladimir

chernishova@arriah.ruФГБУ «Федеральный центр охраны здоровья животных» (ФГБУ «ВНИИЗЖ»)РоссияFGBI “Federal Centre for Animal Health” (FGBI “ARRIAH”)Russian Federation202320092023123197207Copyright © Доронин М.И., Михалишин Д.В., Спрыгин А.В., Мазлум А., Жбанова Т.В., Груздев К.Н., Чернышова Е.В., 20232023Доронин М.И., Михалишин Д.В., Спрыгин А.В., Мазлум А., Жбанова Т.В., Груздев К.Н., Чернышова Е.В.Doronin M.I., Mikhalishin D.V., Sprygin A.V., Mazloum A., Zhbanova T.V., Gruzdev K.N., Chernyshova E.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://veterinary.arriah.ru/jour/article/view/739

В настоящее время в различных областях молекулярной биологии применяется основанная на флуоресцентной детекции количественная полимеразная цепная реакция в режиме реального времени, которая является передовой технологией генной диагностики. Практическая простота, а также сочетание высокой скорости, чувствительности и специфичности сделали возможным использование данного анализа для количественного определения нуклеиновых кислот. В статье представлены общие сведения и отражены рекомендуемые правила для разработки методик количественного ПЦР-анализа в режиме реального времени. Материалы публикации нацелены на предоставление исследователям и рецензентам необходимых требований, которых следует придерживаться, чтобы обеспечить высокую точность, надежность и прозрачность экспериментов, правильную интерпретацию и повторяемость результатов анализа. Представлены современные подходы, которые позволяют получать надежные и достоверные результаты, проводимые разными операторами, в разное время и в разных лабораториях. Приведены основные требования, предъявляемые к применяемым реагентам, перечням нуклеотидных последовательностей и методам проведения валидационного анализа. В целом в представленной публикации отражена информация для достижения трех конечных целей работы: предоставить авторам широкий арсенал инструментов и требований для разработки методик на основе количественной полимеразной цепной реакции в режиме реального времени; дать возможность рецензентам и редакторам оценивать качество представленных материалов статей и методических рекомендаций/указаний в соответствии с требуемыми критериями; получать однородные, сопоставимые и надежные результаты исследований, выполненных с помощью данного метода.

Currently fluorescent quantitative real-time polymerase chain reaction, which is a cutting-edge technology in genetic diagnosis, is used in different areas of molecularbiology. Practicaladvantageofsimplicityaswellascombination of high speed, sensitivityandspecificitymadeitpossibletousethisanalysisfor nucleic acidquantitation. Thepaperpresentsgeneral information andrecommendedrules for thedevelopmentof real-timeqPCR. Thepublication isaimedtoacquaint theresearchersandreviewerswith necessaryrequirementstobefollowedin order toensure high accuracy, reliabilityandtransparencyof theexperiments, correct interpretation andrepeatabilityof thetest results. Currentapproachesaredescribedthatallow obtainingreliableandconsistent resultsbydifferentoperators, at differenttimesandin differentlaboratories. Basicrequirementsforreagentsused, nucleotidesequencesandvalidationmethodsaregiven. In general, thepublication givestheinformation neededtoachievethreeultimategoals: toprovidetheauthorswith abroadrangeoftoolsandrequirementsforthedevelopmentof real-time qPCR based-techniques; togivethepossibilitytothereviewersandeditorsofassessingthequalityofarticlesandguidelines/instructions in accordance with the requiredcriteria; toobtain consistentandreliableresultsof testsperformedusingthis method. 

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