Lanthanide-Complex-Enhanced Bioorthogonal Branched DNA Amplification.

Zhao, F; Guan, Y; Su, F; Du, Z; Wen, S; Zhang, L; Jin, D

Lanthanide-Complex-Enhanced Bioorthogonal Branched DNA Amplification.

Zhao, F Guan, Y Su, F Du, Z Wen, S

Zhang, L

Jin, D

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Publisher:: AMER CHEMICAL SOC
Publication Type:: Journal Article
Citation:: Anal Chem, 2024, 96, (4), pp. 1556-1564
Issue Date:: 2024-01-30

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Field	Value	Language
dc.contributor.author	Zhao, F
dc.contributor.author	Guan, Y
dc.contributor.author	Su, F
dc.contributor.author	Du, Z
dc.contributor.author	Wen, S https://orcid.org/0000-0002-4670-4658
dc.contributor.author	Zhang, L https://orcid.org/0000-0003-0641-2383
dc.contributor.author	Jin, D https://orcid.org/0000-0003-1046-2666
dc.date.accessioned	2024-12-17T00:12:07Z
dc.date.available	2024-12-17T00:12:07Z
dc.date.issued	2024-01-30
dc.identifier.citation	Anal Chem, 2024, 96, (4), pp. 1556-1564
dc.identifier.issn	0003-2700
dc.identifier.issn	1520-6882
dc.identifier.uri	http://hdl.handle.net/10453/182605
dc.description.abstract	Fluorescence in situ hybridization (FISH) is a widely used technique for detecting intracellular nucleic acids. However, its effectiveness in detecting low-copy nucleic acids is limited due to its low fluorescence intensity and background autofluorescence. To address these challenges, we present here an approach of lanthanide-complex-enhanced bioorthogonal-branched DNA amplification (LEBODA) with high sensitivity for in situ nuclear acid detection in single cells. The approach capitalizes on two levels of signal amplification. First, it utilizes click chemistry to directly link a substantial number of bridge probes to target-recognizing probes, providing an initial boost in signal intensity. Second, it incorporates high-density lanthanide complexes into each bridge probe, enabling secondary amplifications. Compared to the traditional "double Z" probes used in the RNAscope method, LEBODA exhibits 4 times the single enhancement for RNA detection signal with the click chemistry approach. Using SARS-CoV-2 pseudovirus-infected HeLa cells, we demonstrate the superiority in the detection of viral-infected cells in rare populations as low as 20% infectious rate. More encouragingly, the LEBODA approach can be adapted for DNA-FISH and single-molecule RNA-FISH, as well as other hybridization-based signal amplification methods. This adaptability broadens the potential applications of LEBODA in the sensitive detection of biomolecules, indicating promising prospects for future research and practical use.
dc.format	Print-Electronic
dc.language	eng
dc.publisher	AMER CHEMICAL SOC
dc.relation	http://purl.org/au-research/grants/arc/FL210100180
dc.relation	http://purl.org/au-research/grants/arc/DE220100846
dc.relation.ispartof	Anal Chem
dc.relation.isbasedon	10.1021/acs.analchem.3c04274
dc.rights	info:eu-repo/semantics/closedAccess
dc.subject	0301 Analytical Chemistry, 0399 Other Chemical Sciences
dc.subject.classification	Analytical Chemistry
dc.subject.classification	3205 Medical biochemistry and metabolomics
dc.subject.classification	3401 Analytical chemistry
dc.subject.classification	4004 Chemical engineering
dc.subject.mesh	Humans
dc.subject.mesh	In Situ Hybridization, Fluorescence
dc.subject.mesh	Lanthanoid Series Elements
dc.subject.mesh	HeLa Cells
dc.subject.mesh	RNA
dc.subject.mesh	DNA
dc.subject.mesh	DNA Probes
dc.subject.mesh	Hela Cells
dc.subject.mesh	Humans
dc.subject.mesh	Lanthanoid Series Elements
dc.subject.mesh	DNA
dc.subject.mesh	DNA Probes
dc.subject.mesh	RNA
dc.subject.mesh	In Situ Hybridization, Fluorescence
dc.subject.mesh	Humans
dc.subject.mesh	In Situ Hybridization, Fluorescence
dc.subject.mesh	Lanthanoid Series Elements
dc.subject.mesh	HeLa Cells
dc.subject.mesh	RNA
dc.subject.mesh	DNA
dc.subject.mesh	DNA Probes
dc.title	Lanthanide-Complex-Enhanced Bioorthogonal Branched DNA Amplification.
dc.type	Journal Article
utslib.citation.volume	96
utslib.location.activity	United States
utslib.for	0301 Analytical Chemistry
utslib.for	0399 Other Chemical Sciences
pubs.organisational-group	University of Technology Sydney
pubs.organisational-group	University of Technology Sydney/Faculty of Science
pubs.organisational-group	University of Technology Sydney/Faculty of Science/School of Mathematical and Physical Sciences
pubs.organisational-group	University of Technology Sydney/UTS Groups
pubs.organisational-group	University of Technology Sydney/UTS Groups/Institute of Biomedical Materials and Devices (IBMD)
utslib.copyright.status	closed_access	*
dc.date.updated	2024-12-17T00:12:05Z
pubs.issue	4
pubs.publication-status	Published
pubs.volume	96
utslib.citation.issue	4

Abstract:

Fluorescence in situ hybridization (FISH) is a widely used technique for detecting intracellular nucleic acids. However, its effectiveness in detecting low-copy nucleic acids is limited due to its low fluorescence intensity and background autofluorescence. To address these challenges, we present here an approach of lanthanide-complex-enhanced bioorthogonal-branched DNA amplification (LEBODA) with high sensitivity for in situ nuclear acid detection in single cells. The approach capitalizes on two levels of signal amplification. First, it utilizes click chemistry to directly link a substantial number of bridge probes to target-recognizing probes, providing an initial boost in signal intensity. Second, it incorporates high-density lanthanide complexes into each bridge probe, enabling secondary amplifications. Compared to the traditional "double Z" probes used in the RNAscope method, LEBODA exhibits 4 times the single enhancement for RNA detection signal with the click chemistry approach. Using SARS-CoV-2 pseudovirus-infected HeLa cells, we demonstrate the superiority in the detection of viral-infected cells in rare populations as low as 20% infectious rate. More encouragingly, the LEBODA approach can be adapted for DNA-FISH and single-molecule RNA-FISH, as well as other hybridization-based signal amplification methods. This adaptability broadens the potential applications of LEBODA in the sensitive detection of biomolecules, indicating promising prospects for future research and practical use.

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http://hdl.handle.net/10453/182605