mimmunМедицинская иммунологияMedical Immunology (Russia)1563-06252313-741XSPb RAACI10.15789/1563-0625-2019-3-527-538mimmun-1826Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИORIGINAL ARTICLESОПРЕДЕЛЕНИЕ РЕФЕРЕНСНЫХ ИНТЕРВАЛОВ TREC И KREC ДЛЯ СКРИНИНГА НОВОРОЖДЕННЫХ С ИММУНОДЕФИЦИТНЫМИ СОСТОЯНИЯМИ В РФDETERMINING REFERENCE RANGES FOR TREC AND KREC ASSAYS IN IMMUNE DEFICIENCY SCREENING OF NEWBORNS IN RUSSIAN FEDERATIONГордуковаМ. А.GordukovaM. A.

биолог клинической диагностической лаборатории

123317, Москва, Шмитовский проезд, 29.Тел.: 8 (499) 256-21-62. Факс: 8 (499) 256-61-27.

Clinical Biologist, Clinical Diagnostic Laboratory

123317, Moscow, Shmitovsky lane, 29.Phone: 7 (499) 256-21-62. Fax: 7 (499) 256-61-27.

ma.gordukova@dgkb-9.ruКорсунскийИ. А.KorsunskyI. A.

к.м.н., заведующий Московским городским центром детской иммунологии и аллергологии

PhD (Medicine), Head, Moscow City Center for Pediatric Immunology and Allergy

ЧурсиноваЮ. В.ChursinovaYu. V.

заведующая отделением организации и сопровождения клинических исследований, ассистент кафедры клинической лабораторной диагностики факультета усовершенствования врачей

г. Москва

Head, Department of Arrangement and Management of Clinical Studies, Assistant Professor, Department of Clinical Laboratory Diagnostics, Faculty of Postgraduate Education

Moscow

БяховаМ. М.ByakhovaM. M.

к.м.н., старший научный сотрудник патологоанатомического отделения

г. Москва

PhD (Medicine), Senior Research Associate, Pathology Department

Moscow

ОскорбинИ. П.OscorbinI. P.

к.б.н., младший научный сотрудник лаборатории фармакогеномики

г. Новосибирск

PhD (Biology), Junior Research Associate, Laboratory of Pharmacogenomics

Novosibirsk

ПродеусА. П.ProdeusA. P.

д.м.н., профессор, заместитель главного врача по инфекционным болезням, Москва;

профессор-консультант, г. Калининград.

PhD, MD (Medicine), Professor, Deputy Medical Director for Infectious Diseases, Moscow;

Consulting Professor, Kaliningrad.

ФилипенкоМ. Л.FilipenkoM. L.

к.б.н., заведующий лабораторией фармакогеномики; заведующий лабораторией змолекулярной диагностики

г. Новосибирск

PhD (Biology), Head, Laboratory of Pharmacogenomics Laboratory; Head, Laboratory of Molecular Ddiagnostics

Novosibirsk

ГБУЗ «Детская городская клиническая больница № 9 имени Г.Н. Сперанского ДЗМ», МоскваРоссияG. Speransky Children’s Hospital No. 9, MoscowRussian FederationГБУЗ «Московский областной научно-исследовательский клинический институт имени М.Ф. Владимирского»РоссияM. Vladimirsky Moscow Regional Research and Clinical Institute (MONIKI)Russian FederationФГБУН «Институт химической биологии и фундаментальной медицины СО РАН»РоссияInstitute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of SciencesRussian FederationГБУЗ «Детская городская клиническая больница № 9 имени Г.Н. Сперанского ДЗМ», Москва; ФГАОУ ВО «Балтийский федеральный университет имени И. Канта»РоссияG. Speransky Children’s Hospital No. 9, Moscow; Immanuel Kant Baltic Federal UniversityRussian FederationФГБУН «Институт химической биологии и фундаментальной медицины СО РАН»; ФГАОУ ВО «Новосибирский национальный исследовательский государственный университет»РоссияInstitute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences; Novosibirsk State University (NSU)Russian Federation201912072019213527538Copyright © Гордукова М.А., Корсунский И.А., Чурсинова Ю.В., Бяхова М.М., Оскорбин И.П., Продеус А.П., Филипенко М.Л., 20192019Гордукова М.А., Корсунский И.А., Чурсинова Ю.В., Бяхова М.М., Оскорбин И.П., Продеус А.П., Филипенко М.Л.Gordukova M.A., Korsunsky I.A., Chursinova Y.V., Byakhova M.M., Oscorbin I.P., Prodeus A.P., Filipenko M.L.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.mimmun.ru/mimmun/article/view/1826

В настоящей работе мы использовали популяционную выборку сухих пятен крови новорожденных на картах Гатри размером 2739 индивидуальных образцов для определения референсного интервала значений молекул ДНК TREC и KREC с целью диагностики первичных иммунодефицитных состояний в неонатальном скрининге. Медиана полученных абсолютных значений TREC и KREC составила 195 (CI95%: 185-206) и 185 (CI95%: 176-197) копий на мкл, соответственно, нормированных значений для TREC – 2780 (CI95%: 2690-2840) и для KREC – 2790 (CI95%: 2700-2900) копий на 2 × 105 копий гена альбумина или 105 ядросодержащих клеток. Референсный интервал рассчитывался для 99 и 99,9 перцентилей всех величин TREC и KREC. Для фильтрации «выпадающих» значений (outliers) был применен критерий Тьюки после логарифмической трансформации данных, ввиду их несимметричного распределения. При анализе абсолютных значений (TREC/KREC на мкл крови) не было идентифицировано «выпадающих» значений TREC, для KREC из дальнейшего анализа было исключено 18 значений (от 9,8 до 13,5). В нормированных значениях TREC/KREC «выбросов» не выявлено. Полученные референсные значения TREC и KREC (на уровне 0,1 перцентиля) составили 458 и 32 на 105 ядросодержащих клеток, 23 и 17 на мкл крови новорожденных соответственно.

In this work, we used a reference population of newborns and sampled dried blood spots on Guthrie cards of 2,739 individual samples to determine the reference intervals for TRECs and KRECs values, in order to diagnose primary immunodeficiency by means of neonatal screening. The median absolute values for TRECs and KRECs were 195 (CI95%: 185-206) and 185 (CI95%: 176-197) copies per μl, respectively; the normalized value for TRECs was 2780 (CI95%: 2690-2840), and for KRECs, 2790 (CI95%: 2700-2900) copies per 2 × 105 copies of the albumin gene or 105 cells. The reference interval was calculated for 99 and 99.9 percentiles of total TRECs and KRECs individual values. Due to asymmetric distribution of data, the outliers were filtered off, using the Tukey’s criterion applied after logarithmic transformation of the data. When analyzing absolute values for TREC/KREC (per μL of blood), no “drop-down” TRECs values were identified; for KRECs, 18 experimental values were excluded from further analysis (from 9.8 to 13.5). The outlying values were not identified among the normalized values of TRECs/KRECs. The obtained reference values for TRECs and KRECs (at the 0.1 percentile level) were, respectively, 458 and 32 per 105 cells, or 23 and 17 per μl of blood samples from neonates.

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