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Comparison of chemiluminescent heterogeneous and homogeneous-heterogeneous assays coupled with isothermal circular strand-displacement polymerization reaction amplification for the quantification of miRNA-141.
- Source :
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The Analyst [Analyst] 2022 Sep 26; Vol. 147 (19), pp. 4293-4300. Date of Electronic Publication: 2022 Sep 26. - Publication Year :
- 2022
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Abstract
- 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 circular strand-displacement polymerization reaction (ICSDPR), which was carried out in heterogeneous and homogeneous media, respectively. In addition, a streptavidin-polyperoxidase conjugate and enhanced chemiluminescence were used to increase the assay's sensitivity. A comparison of the developed assays showed 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-heterogeneous assays were 51 fM and 10 pM, respectively. The amplification index for the ICSDPR used in the heterogeneous assay of miRNA-141 was 100. Using miRNAs of the miRNA-200 family, the high specificity of the assay was demonstrated. MiRNA-141 in human cultured cells was determined by the heterogeneous ICSDPR-assisted assay with chemiluminescence detection. To assess the purification yield of miRNAs from cellular lysates, the heterogeneous assay of miRNA-39 developed on the same platform was used. The intracellular content of miRNA-141 in Caco-2, HepG2, MCF-7 and HeLa was shown to be 3400, 1400, 1300 and 470 copies per cell, respectively.
Details
- Language :
- English
- ISSN :
- 1364-5528
- Volume :
- 147
- Issue :
- 19
- Database :
- MEDLINE
- Journal :
- The Analyst
- Publication Type :
- Academic Journal
- Accession number :
- 36001033
- Full Text :
- https://doi.org/10.1039/d2an00921h