Lanthanide-Complex-Enhanced Bioorthogonal Branched DNA Amplification

Zhao，Fang1; Guan，Yunpeng2; Su，Fei2; Du，Zhongbo1; Wen，Shihui2; Zhang，Le2; Jin，Dayong1,2

2024-01-30

10.1021/acs.analchem.3c04274

Analytical Chemistry   影响因子和分区

0003-2700

1520-6882

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.

SCI ; EI

英语

第一

CHEMISTRY

2-s2.0-85182580628

Scopus

1.Department of Biomedical Engineering,Southern University of Science and Technology,Shenzhen,518055,China
2.Institute for Biomedical Materials and Devices (IBMD),Faculty of Science,University of Technology Sydney,Ultimo,2007,Australia

Zhao，Fang,Guan，Yunpeng,Su，Fei,et al. Lanthanide-Complex-Enhanced Bioorthogonal Branched DNA Amplification[J]. Analytical Chemistry,2024,96(4):1556-1564.

Zhao，Fang.,Guan，Yunpeng.,Su，Fei.,Du，Zhongbo.,Wen，Shihui.,...&Jin，Dayong.(2024).Lanthanide-Complex-Enhanced Bioorthogonal Branched DNA Amplification.Analytical Chemistry,96(4),1556-1564.

Zhao，Fang,et al."Lanthanide-Complex-Enhanced Bioorthogonal Branched DNA Amplification".Analytical Chemistry 96.4(2024):1556-1564.