Recombinase polymerase and loop isothermal amplification in DNA diagnostics of infectious diseases

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Resumo

Рекомбиназная полимеразная и петлевая изотермическая амплификация может быть проведена во внелабораторных условиях, что делает эти методы перспективными для разработки экспресс-тестов для ДНК-диагностики инфекционных заболеваний человека, а также сельскохозяйственных животных и растений в формате оказания медицинской помощи на месте (англ. “point-of-care” testing) или полевой детекции (англ. “in-field” detection). В обзоре рассмотрены основные принципы, на которых основаны данные методы, а также их современное состояние с акцентом на неинструментальные методы регистрации результата изотермической амплификации с помощью колориметрии и иммунохроматографических тест-полосок. Подробно освещены подходы к повышению селективности изотермической амплификации путём её объединения с CRISPR/Cas-детекцией или путём объединения двух методов по принципу гнездовой амплификации. 

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Sobre autores

L. Kurbatov

Научно-исследовательский институт биомедицинской химии имени В.Н. Ореховича

Email: lenasuprun@mail.ru
Rússia, ул. Погодинская, 10, стр. 8, Москва, 119121

K. Ptitsyn

Научно-исследовательский институт биомедицинской химии имени В.Н. Ореховича

Email: lenasuprun@mail.ru
Rússia, ул. Погодинская, 10, стр. 8, Москва, 119121

S. Khmeleva

Научно-исследовательский институт биомедицинской химии имени В.Н. Ореховича

Email: lenasuprun@mail.ru
Rússia, ул. Погодинская, 10, стр. 8, Москва, 119121

S. Radko

Научно-исследовательский институт биомедицинской химии имени В.Н. Ореховича

Email: lenasuprun@mail.ru
Rússia, ул. Погодинская, 10, стр. 8, Москва, 119121

A. Lisitsa

Научно-исследовательский институт биомедицинской химии имени В.Н. Ореховича; Тюменский государственный университет

Email: enasuprun@mail.ru
Rússia, ул. Погодинская, 10, стр. 8, Москва, 119121; ул. Володарского, 6, Тюмень, 625003

E. Suprun

Научно-исследовательский институт биомедицинской химии имени В.Н. Ореховича; Московский государственный университет имени М.В. Ломоносова

Autor responsável pela correspondência
Email: lenasuprun@mail.ru

химический факультет

Rússia, ул. Погодинская, 10, стр. 8, Москва, 119121; Ленинские горы, 1, стр. 3, ГСП-1, Москва, 119991

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2. Figure 1. Schematic representation of recombinant polymerase amplification (RPA).

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3. Fig. 2. Schematic representation of loop amplification (LAMP).

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4. Pain. 3. Schematic representation of the detection of PRO and LAMP amplicons using CRISPR/Cas nucleases Cas13a and Cas 12 (Cas12a or Cas12b).

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