High-Throughput Detection of Multiple miRNAs and Methylated DNA by Droplet Digital PCR
Altered miRNA expression and DNA methylation have highly active and diverse roles in carcinogenesis. Simultaneous detection of the molecular aberrations may have a synergistic effect on the diagnosis of malignancies. Herein, we develop a high-throughput assay for detecting multiple miRNAs and DNA me...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-04-01
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| Series: | Journal of Personalized Medicine |
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| Online Access: | https://www.mdpi.com/2075-4426/11/5/359 |
| _version_ | 1797535768700256256 |
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| author | Ning Li Pushpa Dhilipkannah Feng Jiang |
| author_facet | Ning Li Pushpa Dhilipkannah Feng Jiang |
| author_sort | Ning Li |
| collection | DOAJ |
| description | Altered miRNA expression and DNA methylation have highly active and diverse roles in carcinogenesis. Simultaneous detection of the molecular aberrations may have a synergistic effect on the diagnosis of malignancies. Herein, we develop a high-throughput assay for detecting multiple miRNAs and DNA methylation using droplet digital PCR (ddPCR) coupled with a 96-microwell plate. The microplate-based ddPCR could absolutely and reproducibly quantify 15 miRNAs and 14 DNA methylation sites with a high sensitivity (one copy/µL and 0.1%, respectively). Analyzing sputum and plasma of 40 lung cancer patients and 36 cancer-free smokers by this approach identified an integrated biomarker panel consisting of two sputum miRNAs (miRs-31-5p and 210-3p), one sputum DNA methylation (RASSF1A), and two plasma miRNAs (miR-21-5p and 126) for the diagnosis of lung cancer with higher sensitivity and specificity compared with a single type of biomarker. The diagnostic value of the integrated biomarker panel for the early detection of lung cancer was confirmed in a different cohort of 36 lung cancer patients and 39 cancer-free smokers. The high-throughput assay for quantification of multiple molecular aberrations across sputum and plasma could improve the early detection of lung cancer. |
| first_indexed | 2024-03-10T11:50:01Z |
| format | Article |
| id | doaj.art-3ea3fac3760b4a128467be1929d8db55 |
| institution | Directory Open Access Journal |
| issn | 2075-4426 |
| language | English |
| last_indexed | 2024-03-10T11:50:01Z |
| publishDate | 2021-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Journal of Personalized Medicine |
| spelling | doaj.art-3ea3fac3760b4a128467be1929d8db552023-11-21T17:48:27ZengMDPI AGJournal of Personalized Medicine2075-44262021-04-0111535910.3390/jpm11050359High-Throughput Detection of Multiple miRNAs and Methylated DNA by Droplet Digital PCRNing Li0Pushpa Dhilipkannah1Feng Jiang2Department of Pathology, University of Maryland School of Medicine, 10 S. Pine St., Baltimore, MD 21201, USADepartment of Pathology, University of Maryland School of Medicine, 10 S. Pine St., Baltimore, MD 21201, USADepartment of Pathology, University of Maryland School of Medicine, 10 S. Pine St., Baltimore, MD 21201, USAAltered miRNA expression and DNA methylation have highly active and diverse roles in carcinogenesis. Simultaneous detection of the molecular aberrations may have a synergistic effect on the diagnosis of malignancies. Herein, we develop a high-throughput assay for detecting multiple miRNAs and DNA methylation using droplet digital PCR (ddPCR) coupled with a 96-microwell plate. The microplate-based ddPCR could absolutely and reproducibly quantify 15 miRNAs and 14 DNA methylation sites with a high sensitivity (one copy/µL and 0.1%, respectively). Analyzing sputum and plasma of 40 lung cancer patients and 36 cancer-free smokers by this approach identified an integrated biomarker panel consisting of two sputum miRNAs (miRs-31-5p and 210-3p), one sputum DNA methylation (RASSF1A), and two plasma miRNAs (miR-21-5p and 126) for the diagnosis of lung cancer with higher sensitivity and specificity compared with a single type of biomarker. The diagnostic value of the integrated biomarker panel for the early detection of lung cancer was confirmed in a different cohort of 36 lung cancer patients and 39 cancer-free smokers. The high-throughput assay for quantification of multiple molecular aberrations across sputum and plasma could improve the early detection of lung cancer.https://www.mdpi.com/2075-4426/11/5/359miRNAmethylationsputumplasmalung cancer |
| spellingShingle | Ning Li Pushpa Dhilipkannah Feng Jiang High-Throughput Detection of Multiple miRNAs and Methylated DNA by Droplet Digital PCR Journal of Personalized Medicine miRNA methylation sputum plasma lung cancer |
| title | High-Throughput Detection of Multiple miRNAs and Methylated DNA by Droplet Digital PCR |
| title_full | High-Throughput Detection of Multiple miRNAs and Methylated DNA by Droplet Digital PCR |
| title_fullStr | High-Throughput Detection of Multiple miRNAs and Methylated DNA by Droplet Digital PCR |
| title_full_unstemmed | High-Throughput Detection of Multiple miRNAs and Methylated DNA by Droplet Digital PCR |
| title_short | High-Throughput Detection of Multiple miRNAs and Methylated DNA by Droplet Digital PCR |
| title_sort | high throughput detection of multiple mirnas and methylated dna by droplet digital pcr |
| topic | miRNA methylation sputum plasma lung cancer |
| url | https://www.mdpi.com/2075-4426/11/5/359 |
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