Comparison of chemiluminescent heterogeneous and homogeneous–heterogeneous assays coupled with isothermal circular strand-displacement polymerization reaction amplification for the quantification of miRNA-141

Solovjev, A.M.; Galkin, I.I.; Medved’ko, A.V.; Pletjushkina, O.Y.; Zhao, S.; Sakharov, I.Y.

Авторы: Solovjev Anton M., Galkin Ivan I., Medved’ko Alexey V., Pletjushkina Olga Yu, Zhao Shulin, Sakharov Ivan Yu
Журнал: The Analyst
Номер: 147
Год издания: 2022
Издательство: Royal Society of Chemistry
Местоположение издательства: United Kingdom
Первая страница: 4293
Последняя страница: 4300
DOI: 10.1039/d2an00921h
Аннотация: Heterogeneous and homogeneous–heterogeneous assays for the quantitation of hsa-miR-141-3p(miRNA-141) were constructed. Both microplate assays were based on the use of the isothermal circularstrand-displacement polymerization reaction (ICSDPR), which was carried out in heterogeneous andhomogeneous media, respectively. In addition, a streptavidin-polyperoxidase conjugate and enhancedchemiluminescence were used to increase the assay’s sensitivity. A comparison of the developed assaysshowed that the sensitivity of the heterogeneous assay was higher than that of the homogeneous–heterogeneous assay. The detection limit values of the heterogeneous and homogeneous–heterogeneousassays were 51 fM and 10 pM, respectively. The amplification index for the ICSDPR used in theheterogeneous assay of miRNA-141 was 100. Using miRNAs of the miRNA-200 family, the high specificityof the assay was demonstrated. MiRNA-141 in human cultured cells was determined by the heterogeneousICSDPR-assisted assay with chemiluminescence detection. To assess the purification yield ofmiRNAs from cellular lysates, the heterogeneous assay of miRNA-39 developed on the same platformwas used. The intracellular content of miRNA-141 in Caco-2, HepG2, MCF-7 and HeLa was shown to be3400, 1400, 1300 and 470 copies per cell, respectively.
Добавил в систему: Сахаров Иван Юрьевич

	ИСТИНА	Войти в систему Регистрация
	ФНКЦ РР
	Главная Поиск Статистика О проекте Помощь

ИСТИНА

ФНКЦ РР

Comparison of chemiluminescent heterogeneous and homogeneous–heterogeneous assays coupled with isothermal circular strand-displacement polymerization reaction amplification for the quantification of miRNA-141статья Исследовательская статья

Прикрепленные файлы