МЕТОДЫ ДЕТЕКЦИИ РЕЗУЛЬТАТОВ РЕАКЦИИ ПЕТЛЕВОЙ ИЗОТЕРМИЧЕСКОЙ АМПЛИФИКАЦИИ ДНК

Аннотация

Метод петлевой изотермической амплификации ДНК (LAMP) был разработан T. Notomi с соавт. в 2000 г. и, благодаря своей точности, чувствительности и простоте исполнения, стал одним из самых перспективных методов для осуществления диагностики на месте лечения («point-of-care»). В настоящем обзоре рассмотрены разные способы детекции результатов реакции LAMP, выявлены их преимущества и недостатки. Методы детекции результатов LAMP могут быть условно подразделены на косвенные и прямые. К косвенным методам, направленным на выявление изменений в химическом составе реакционной смеси, можно отнести турбидиметрию в реальном времени, флуоресцентную детекцию с кальцеином, колориметрическую детекцию с гидроксинафтоловым синим и детекцию с использованием модифицированных наночастиц золота. Прямые методы, основанные на выявлении накопления в процессе реакции ампликонов, включают в себя флуориметрическое выявление ампликонов с применением интеркалирующих красителей, резонансный перенос энергии флуоресценции, иммуноферментный анализ, реакцию иммунохроматографии, использование катионных полимеров и наночастиц золота. Для разработок в области диагностики «point-of-care» характерна выраженная тенденция к миниатюризации, проведению реакции LAMP на микрочипах и микрофлюидных устройствах с электрохимическим или оптическим способом детекции. Наиболее перспективными для диагностики инфекционных заболеваний являются методы турбидиметрии и использование интеркалирующих красителей. Разработка отечественных портативных приборов для детекции результатов LAMP, основанных на флуоресцентной детекции или турбидиметрии в реальном времени будет способствовать широкому внедрению метода в практику клинической лабораторной диагностики. Поиск литературы проводился в базе данный Pubmed ncbi по ключевым словам.

Об авторах

Список литературы

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