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Simultaneous detection of dual microRNAs related to EV71 using ICP-MS based on metal nanoparticle labeling with
Yuxin Wang1, Lijun Shao2, Zhigang Zhao2
1Department of Physical and Chemical Inspection, School of Public Health, Cheeloo College of Medicine, Shandong University, Jinan, PR China; Shandong Academy of Preventive Medicine, Shandong Center for Food Safety Risk Assessment, Shandong Center for Disease Control and Prevention, Jinan, PR China; Department of Transfusion Medicine, West China Hospital, Sichuan University, Chengdu, PR China.
Background:
Hand, foot, and mouth (HMFD) disease caused by enterovirus 71 (EV 71), is closely associated with severe clinical manifestations and can be deadly. Early detection of EV 71 can be achieved by detecting the increment in miR296 and miR16 in the serum. Using HCR to amplify signals and convert biological signals into metal nanoparticle signals detectable by ICP-MS is a detection method that can collect more accurate and reliable information, compared with traditional methods, in the detection of biological samples.
Results:
We described a strategy for the simultaneous detection of miR296 and miR16 by ICP-MS based on metal nanoparticles (NPs) labeling with HCR. Briefly, single-stranded DNA (ssDNA) and magnetic beads (MBs), as well as NPs and signal probes for miRNA (Sp-miR) were firstly conjugated via the streptavidin-biotin recognition system, constituting ssDNA-MBs and NPs-Sp-miR complex, respectively. The latter complex then hybridized with the former through HCR, generating the nanosensors for targets. Then, the targets were added and hybridized with ssDNA, and the HCR complex with NPs was released into the solution. Finally, the corresponding signals of the NPs were measured by ICP-MS. Results demonstrated that the developed method had good sensitivity and satisfactory selectivity and precision. Furthermore, when applied to biological samples with a complex matrix, the developed method also showed good recovery (88 % - 92 %) and reproducibility (RSD<10 %).
Significance:
This method contributes to the early diagnosis of HFMD and opens up ideas for the further development of high-throughput biomarker detection. The strategy has practical potential for miR296 and miR16 detection in biological samples and provides a promising tool for multiple miRNA detection.
Insights
Early detection of hand, foot, and mouth disease (HFMD) is possible by measuring miR296 and miR16 levels. A new ICP-MS method using HCR and nanoparticles offers accurate and sensitive detection in biological samples.
Area of Science:
- Biomedical Engineering
- Analytical Chemistry
- Molecular Diagnostics
Background:
- Hand, foot, and mouth disease (HFMD) caused by enterovirus 71 (EV71) can lead to severe, fatal outcomes.
- Early diagnosis of EV71 infection is crucial and can be achieved by monitoring serum miR296 and miR16 levels.
- Traditional detection methods may lack accuracy and reliability for biological samples.
Purpose of the Study:
- To develop a sensitive and selective method for simultaneous detection of miR296 and miR16.
- To utilize hybridization chain reaction (HCR) and inductively coupled plasma mass spectrometry (ICP-MS) for enhanced signal amplification and detection.
- To provide a reliable tool for early HFMD diagnosis and high-throughput biomarker detection.
Main Methods:
- A novel strategy involving metal nanoparticle (NP) labeling and HCR amplification was employed.
- Single-stranded DNA (ssDNA) and NPs were conjugated to magnetic beads (MBs) and signal probes (Sp-miR), respectively.
- Nanosensors were formed via HCR hybridization, followed by target binding, NP release, and ICP-MS detection.
Main Results:
- The developed method demonstrated good sensitivity, selectivity, and precision for detecting miR296 and miR16.
- The method achieved high recovery rates (88%-92%) and reproducibility (RSD<10%) in complex biological matrices.
- This approach offers a significant improvement over traditional detection techniques.
Conclusions:
- The developed ICP-MS based method enables early diagnosis of HFMD.
- This strategy provides a promising tool for multiple miRNA detection in biological samples.
- The approach has potential for high-throughput biomarker analysis and future diagnostic applications.
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