Cargando…

Simultaneous and ultrasensitive detection of multiple microRNAs by single-molecule fluorescence imaging

Cell status changes are typically accompanied by the simultaneous changes of multiple microRNA (miRNA) levels. Thus, simultaneous and ultrasensitive detection of multiple miRNA biomarkers shows great promise in early cancer diagnosis. Herein, a facile single-molecule fluorescence imaging assay was p...

Descripción completa

Detalles Bibliográficos
Autores principales: Zhang, Hongding, Huang, Xuedong, Liu, Jianwei, Liu, Baohong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Royal Society of Chemistry 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8152581/
https://www.ncbi.nlm.nih.gov/pubmed/34122849
http://dx.doi.org/10.1039/d0sc00580k
_version_ 1783698625795194880
author Zhang, Hongding
Huang, Xuedong
Liu, Jianwei
Liu, Baohong
author_facet Zhang, Hongding
Huang, Xuedong
Liu, Jianwei
Liu, Baohong
author_sort Zhang, Hongding
collection PubMed
description Cell status changes are typically accompanied by the simultaneous changes of multiple microRNA (miRNA) levels. Thus, simultaneous and ultrasensitive detection of multiple miRNA biomarkers shows great promise in early cancer diagnosis. Herein, a facile single-molecule fluorescence imaging assay was proposed for the simultaneous and ultrasensitive detection of multiple miRNAs using only one capture anti-DNA/RNA antibody (S9.6 antibody). Two complementary DNAs (cDNAs) designed to hybridize with miRNA-21 and miRNA-122 were labelled with Cy3 (cDNA1) and Cy5 (cDNA2) dyes at their 5′-ends, respectively. After hybridization, both miRNA-21/cDNA1 and miRNA-122/cDNA2 complexes were captured by S9.6 antibodies pre-modified on a coverslip surface. Subsequently, the Cy3 and Cy5 dyes on the coverslip surface were imaged by the single-molecule fluorescence setup. The amount of miRNA-21 and miRNA-122 was quantified by counting the image spots from the Cy3 and Cy5 dye molecules in the green and red channels, respectively. The proposed assay displayed high specificity and sensitivity for singlet miRNA detection both with a detection limit of 5 fM and for multiple miRNA detection both with a detection limit of 20 fM. Moreover, it was also demonstrated that the assay could be used to detect multiple miRNAs simultaneously in human hepatocellular cancer cells (HepG2 cells). The proposed assay provides a novel biosensing platform for the ultrasensitive and simple detection of multiple miRNA expressions and shows great prospects for early cancer diagnosis.
format Online
Article
Text
id pubmed-8152581
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher The Royal Society of Chemistry
record_format MEDLINE/PubMed
spelling pubmed-81525812021-06-11 Simultaneous and ultrasensitive detection of multiple microRNAs by single-molecule fluorescence imaging Zhang, Hongding Huang, Xuedong Liu, Jianwei Liu, Baohong Chem Sci Chemistry Cell status changes are typically accompanied by the simultaneous changes of multiple microRNA (miRNA) levels. Thus, simultaneous and ultrasensitive detection of multiple miRNA biomarkers shows great promise in early cancer diagnosis. Herein, a facile single-molecule fluorescence imaging assay was proposed for the simultaneous and ultrasensitive detection of multiple miRNAs using only one capture anti-DNA/RNA antibody (S9.6 antibody). Two complementary DNAs (cDNAs) designed to hybridize with miRNA-21 and miRNA-122 were labelled with Cy3 (cDNA1) and Cy5 (cDNA2) dyes at their 5′-ends, respectively. After hybridization, both miRNA-21/cDNA1 and miRNA-122/cDNA2 complexes were captured by S9.6 antibodies pre-modified on a coverslip surface. Subsequently, the Cy3 and Cy5 dyes on the coverslip surface were imaged by the single-molecule fluorescence setup. The amount of miRNA-21 and miRNA-122 was quantified by counting the image spots from the Cy3 and Cy5 dye molecules in the green and red channels, respectively. The proposed assay displayed high specificity and sensitivity for singlet miRNA detection both with a detection limit of 5 fM and for multiple miRNA detection both with a detection limit of 20 fM. Moreover, it was also demonstrated that the assay could be used to detect multiple miRNAs simultaneously in human hepatocellular cancer cells (HepG2 cells). The proposed assay provides a novel biosensing platform for the ultrasensitive and simple detection of multiple miRNA expressions and shows great prospects for early cancer diagnosis. The Royal Society of Chemistry 2020-03-24 /pmc/articles/PMC8152581/ /pubmed/34122849 http://dx.doi.org/10.1039/d0sc00580k Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by-nc/3.0/
spellingShingle Chemistry
Zhang, Hongding
Huang, Xuedong
Liu, Jianwei
Liu, Baohong
Simultaneous and ultrasensitive detection of multiple microRNAs by single-molecule fluorescence imaging
title Simultaneous and ultrasensitive detection of multiple microRNAs by single-molecule fluorescence imaging
title_full Simultaneous and ultrasensitive detection of multiple microRNAs by single-molecule fluorescence imaging
title_fullStr Simultaneous and ultrasensitive detection of multiple microRNAs by single-molecule fluorescence imaging
title_full_unstemmed Simultaneous and ultrasensitive detection of multiple microRNAs by single-molecule fluorescence imaging
title_short Simultaneous and ultrasensitive detection of multiple microRNAs by single-molecule fluorescence imaging
title_sort simultaneous and ultrasensitive detection of multiple micrornas by single-molecule fluorescence imaging
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8152581/
https://www.ncbi.nlm.nih.gov/pubmed/34122849
http://dx.doi.org/10.1039/d0sc00580k
work_keys_str_mv AT zhanghongding simultaneousandultrasensitivedetectionofmultiplemicrornasbysinglemoleculefluorescenceimaging
AT huangxuedong simultaneousandultrasensitivedetectionofmultiplemicrornasbysinglemoleculefluorescenceimaging
AT liujianwei simultaneousandultrasensitivedetectionofmultiplemicrornasbysinglemoleculefluorescenceimaging
AT liubaohong simultaneousandultrasensitivedetectionofmultiplemicrornasbysinglemoleculefluorescenceimaging