Use of Droplet Digital Polymerase Chain Reaction to Identify Biomarkers for Differentiation of Benign and Malignant Renal Masses
Several microRNAs (miRNAs) have been identified as cell-free biomarkers for detecting renal cell carcinoma (RCC). Droplet digital polymerase chain reaction (ddPCR) is a unique technology for nucleic acid quantification. It has the potential for superior precision, reproducibility, and diagnostic per...
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| Format: | Article |
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MDPI AG
2024-02-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/16/4/787 |
| _version_ | 1797298615261069312 |
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| author | Joshua P. Hayden Adam Wiggins Travis Sullivan Thomas Kalantzakos Kailey Hooper Alireza Moinzadeh Kimberly Rieger-Christ |
| author_facet | Joshua P. Hayden Adam Wiggins Travis Sullivan Thomas Kalantzakos Kailey Hooper Alireza Moinzadeh Kimberly Rieger-Christ |
| author_sort | Joshua P. Hayden |
| collection | DOAJ |
| description | Several microRNAs (miRNAs) have been identified as cell-free biomarkers for detecting renal cell carcinoma (RCC). Droplet digital polymerase chain reaction (ddPCR) is a unique technology for nucleic acid quantification. It has the potential for superior precision, reproducibility, and diagnostic performance in identifying circulating miRNA biomarkers compared to conventional quantitative real-time PCR (qRT-PCR). This study aims to evaluate the performance of ddPCR compared to qRT- PCR in identifying miRNA biomarkers that differentiate malignant from benign renal masses. Potential biomarkers of RCC were identified from a literature review. RNA was extracted from the plasma of 56 patients. All the samples underwent analysis via ddPCR as well as qRT-PCR, and expression levels were recorded for the following miRNAs: miR-93, -144, -210, -221, and -222. Tumors were grouped into low-grade ccRCC, high-grade ccRCC, papillary RCC, and benign masses (primarily angiomyolipoma). The miRNA miR-210 (<i>p</i> = 0.034) and the combination of miRs-210 and miR-222 (<i>p</i> = 0.003) were expressed at significantly higher rates among those with RCC than those with benign masses, as measured by ddPCR. Using the combination of miR-210 and miR-222, ddPCR identified significant differences between the subgroups: papillary RCC versus benign (<i>p</i> = 0.03), low-grade ccRCC versus benign (<i>p</i> = 0.026), and high-grade ccRCC versus benign (<i>p</i> = 0.002). The only significant difference between these subgroups using qRT-PCR was between high-grade ccRCC and benign (<i>p</i> = 0.045). All the AUCs were significant when comparing each RCC subgroup with benign for both PCR technologies. Using a combination of miR-210 and miR-222, ddPCR identified significant differences between benign and malignant renal masses that were not identified as significant by conventional qRT-PCR. |
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| format | Article |
| id | doaj.art-5662a196aa3a466f9f9a34cca1768f9c |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-07T22:38:34Z |
| publishDate | 2024-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cancers |
| spelling | doaj.art-5662a196aa3a466f9f9a34cca1768f9c2024-02-23T15:11:14ZengMDPI AGCancers2072-66942024-02-0116478710.3390/cancers16040787Use of Droplet Digital Polymerase Chain Reaction to Identify Biomarkers for Differentiation of Benign and Malignant Renal MassesJoshua P. Hayden0Adam Wiggins1Travis Sullivan2Thomas Kalantzakos3Kailey Hooper4Alireza Moinzadeh5Kimberly Rieger-Christ6Department of Urology, Lahey Hospital & Medical Center, Burlington, MA 01805, USADepartment of Urology, Lahey Hospital & Medical Center, Burlington, MA 01805, USADepartment of Translational Research, Lahey Hospital & Medical Center, Burlington, MA 01805, USADepartment of Translational Research, Lahey Hospital & Medical Center, Burlington, MA 01805, USADepartment of Translational Research, Lahey Hospital & Medical Center, Burlington, MA 01805, USADepartment of Urology, Lahey Hospital & Medical Center, Burlington, MA 01805, USADepartment of Urology, Lahey Hospital & Medical Center, Burlington, MA 01805, USASeveral microRNAs (miRNAs) have been identified as cell-free biomarkers for detecting renal cell carcinoma (RCC). Droplet digital polymerase chain reaction (ddPCR) is a unique technology for nucleic acid quantification. It has the potential for superior precision, reproducibility, and diagnostic performance in identifying circulating miRNA biomarkers compared to conventional quantitative real-time PCR (qRT-PCR). This study aims to evaluate the performance of ddPCR compared to qRT- PCR in identifying miRNA biomarkers that differentiate malignant from benign renal masses. Potential biomarkers of RCC were identified from a literature review. RNA was extracted from the plasma of 56 patients. All the samples underwent analysis via ddPCR as well as qRT-PCR, and expression levels were recorded for the following miRNAs: miR-93, -144, -210, -221, and -222. Tumors were grouped into low-grade ccRCC, high-grade ccRCC, papillary RCC, and benign masses (primarily angiomyolipoma). The miRNA miR-210 (<i>p</i> = 0.034) and the combination of miRs-210 and miR-222 (<i>p</i> = 0.003) were expressed at significantly higher rates among those with RCC than those with benign masses, as measured by ddPCR. Using the combination of miR-210 and miR-222, ddPCR identified significant differences between the subgroups: papillary RCC versus benign (<i>p</i> = 0.03), low-grade ccRCC versus benign (<i>p</i> = 0.026), and high-grade ccRCC versus benign (<i>p</i> = 0.002). The only significant difference between these subgroups using qRT-PCR was between high-grade ccRCC and benign (<i>p</i> = 0.045). All the AUCs were significant when comparing each RCC subgroup with benign for both PCR technologies. Using a combination of miR-210 and miR-222, ddPCR identified significant differences between benign and malignant renal masses that were not identified as significant by conventional qRT-PCR.https://www.mdpi.com/2072-6694/16/4/787microRNAddPCRrenal cell carcinomacell-free biomarkerbenign |
| spellingShingle | Joshua P. Hayden Adam Wiggins Travis Sullivan Thomas Kalantzakos Kailey Hooper Alireza Moinzadeh Kimberly Rieger-Christ Use of Droplet Digital Polymerase Chain Reaction to Identify Biomarkers for Differentiation of Benign and Malignant Renal Masses Cancers microRNA ddPCR renal cell carcinoma cell-free biomarker benign |
| title | Use of Droplet Digital Polymerase Chain Reaction to Identify Biomarkers for Differentiation of Benign and Malignant Renal Masses |
| title_full | Use of Droplet Digital Polymerase Chain Reaction to Identify Biomarkers for Differentiation of Benign and Malignant Renal Masses |
| title_fullStr | Use of Droplet Digital Polymerase Chain Reaction to Identify Biomarkers for Differentiation of Benign and Malignant Renal Masses |
| title_full_unstemmed | Use of Droplet Digital Polymerase Chain Reaction to Identify Biomarkers for Differentiation of Benign and Malignant Renal Masses |
| title_short | Use of Droplet Digital Polymerase Chain Reaction to Identify Biomarkers for Differentiation of Benign and Malignant Renal Masses |
| title_sort | use of droplet digital polymerase chain reaction to identify biomarkers for differentiation of benign and malignant renal masses |
| topic | microRNA ddPCR renal cell carcinoma cell-free biomarker benign |
| url | https://www.mdpi.com/2072-6694/16/4/787 |
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