ФБУН «Центральный научно-исследовательский институт эпидемиологии» Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человека (Роспотребнадзор), 111123, Москва, Россия В настоящем обзоре представлены основные принципы применения реакции петлевой изотермической амплификации (loop-mediated isothermal amplification, LAMP) для экспресс-диагностики коронавирусной инфекции, вызванной SARS-CoV-2. Кратко описаны базовые технические детали метода, наиболее популярные способы специфической и неспецифической детекции продуктов амплификации, обсуждены первые опубликованные работы по использованию рассматриваемой технологии для выявления фрагментов молекулы нуклеиновой кислоты вируса SARS-CoV-2, в том числе разрабатываемые в Российской Федерации. Для доступных тестов на базе LAMP перечислены основные аналитические характеристики наборов, которые нередко сравнимы с параметрами тест-систем на основе метода полимеразной цепной реакции с обратной транскрипцией (ОТ-ПЦР), а в ряде случаев превосходят их. Обсуждены преимущества и ограничения этого подхода в сравнении с другими способами молекулярной диагностики (в первую очередь ОТ-ПЦР), а также перспективы развития технологии для выявления возбудителей других инфекций.
Introduction. Currently, the basis for molecular diagnostics of most infections is the use of reverse transcription polymerase chain reaction (RT-PCR). Technologies based on reverse transcription isothermal loop amplification (RT-LAMP) can be used as an alternative to RT-PCR for diagnostic purposes. In this study, we compared the RTLAMP and RT-PCR methods in order to analyze both the advantages and disadvantages of the two approaches.Material and methods. For the study, we used reagent kits based on RT-PCR and RT-LAMP. The biological material obtained by taking swabs from the mucous membrane of the oropharynx and nasopharynx in patients with symptoms of a new coronavirus infection was used.Results. We tested 381 RNA samples of the SARS-CoV-2 virus (Coronaviridae: Coronavirinae: Betacoronavirus; Sarbecovirus) from various patients. The obtained values of the threshold cycle (Ct) for RT-PCR averaged 20.0 ± 3.7 s (1530 ± 300 s), and for RT-LAMP 12.8 ± 3.7 s (550 ± 160 s). Proceeding from the theoretical assumptions, a linear relationship between values obtained in two kits was proposed as a hypothesis; the correlation coefficient was approximately 0.827. At the same time, for samples with a low viral load (VL), the higher Ct values in RT-LAMP did not always correlated with those obtained in RT-PCR.Discussion. We noted a significant gain in time for analysis using RT-LAMP compared to RT-PCR, which can be important in the context of testing a large number of samples. Being easy to use and boasting short turnaround time, RT-LAMP-based test systems can be used for mass screening in order to identify persons with medium and high VLs who pose the greatest threat of the spread of SARS-CoV-2, while RT-PCR-based diagnostic methods are also suitable for estimation of VL and its dynamics in patients with COVID-19.
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