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Updated: Oct 17, 2025
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Multiplexed Isothermal Amplification Based Diagnostic Platform to Detect Zika, Chikungunya, and Dengue 1
Published on: March 13, 2018
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Rolling Circle Amplification in Integrated Microsystems: An Uncut Gem toward Massively Multiplexed Pathogen
Ruben R G Soares1,2, Narayanan Madaboosi1,3, Mats Nilsson1
1Department of Biochemistry and Biophysics, Science for Life Laboratory, Stockholm University, 17165 Solna, Sweden.
Accounts of Chemical Research
|October 12, 2021
Summary
Padlock probes (PLPs) combined with rolling circle amplification (RCA) offer a powerful isothermal method for precise nucleic acid detection. This PLP-RCA technology enables highly multiplexed pathogen diagnostics, ideal for point-of-care applications and spatial multiomics.
Area of Science:
- Biotechnology and Biomedical Engineering
- Molecular Diagnostics
- Nucleic Acid Amplification
Background:
- Precise nucleic acid detection is crucial for understanding diseases, developing therapeutics, and improving biotechnological processes.
- The SARS-CoV-2 pandemic highlighted the need for rapid and accurate pathogen diagnostics.
- Current gold standard methods like RT-PCR require thermal cycling, limiting point-of-care applications, and lack high-level multiplexing capabilities.
Purpose of the Study:
- To advocate for padlock probes (PLPs) coupled with rolling circle amplification (RCA), termed PLP-RCA, as an underexploited isothermal nucleic acid amplification test (NAAT).
- To highlight PLP-RCA's potential for unprecedented multiplexing, specificity, versatility, and integration into miniaturized point-of-care (PoC) platforms.
- To explore PLP-RCA's utility in spatial multiomics for pathogenesis research.
Main Methods:
- Detailed explanation of PLP design and ligation mechanisms.
- Description of rolling circle amplification (RCA) following probe ligation.
- Overview of detection methods for RCA products.
- Discussion of PLP-RCA miniaturization and integration strategies for PoC applications.
Main Results:
- PLP-RCA demonstrates high specificity and multiplexing capabilities for detecting viral, bacterial, and fungal pathogens.
- Successful application in genotyping antibiotic resistance genes and viral subtyping.
- Advancements in miniaturization, automation, and signal transduction enable rapid, user-friendly PoC diagnostics.
- PLP-RCA retains spatial information, enabling spatial multiomics analysis.
Conclusions:
- PLP-RCA is a versatile and powerful isothermal NAAT with significant potential for ubiquitous, precise, and unbiased pathogen diagnostics.
- The technology is amenable to integration into miniaturized PoC devices, expediting results and minimizing user input.
- PLP-RCA offers unique advantages for tackling emergent threats like viral variants and antimicrobial resistance.
- Its capacity for spatial multiomics opens new avenues in pathogenesis research.
