Melting curve analyses in the quantitative real-time polymerase chain reaction of methylated/non-methylated DNA toward the detection of oral cancer using gargle fluid
The early detection and early treatment of oral cancer are of paramount importance for improving the patients' prognoses. Our group previously reported a noninvasive test for oral cancer that uses a commercially available methylation-specific multiple ligation-dependent probe amplification (MS-...
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Elsevier
2025-02-01
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405844025006668 |
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| author | Yusaku Noma Hayate Sawayama Kouta Yamashiro Keitaro Nishi Haruka Yoshinaga Ryota Takaoka Tsuyoshi Sugiura Tomonori Sasahira Tomofumi Hamada Yasuo Suda Tatsuo Okui |
| author_facet | Yusaku Noma Hayate Sawayama Kouta Yamashiro Keitaro Nishi Haruka Yoshinaga Ryota Takaoka Tsuyoshi Sugiura Tomonori Sasahira Tomofumi Hamada Yasuo Suda Tatsuo Okui |
| author_sort | Yusaku Noma |
| collection | DOAJ |
| description | The early detection and early treatment of oral cancer are of paramount importance for improving the patients' prognoses. Our group previously reported a noninvasive test for oral cancer that uses a commercially available methylation-specific multiple ligation-dependent probe amplification (MS-MLPA) method, focusing on the DNA methylation of cells in gargle fluid. However, the conventional MS-MLPA method is complicated, costly, and thus difficult to use for screening and other health examinations. We therefore sought to develop a simple screening method for the diagnosis of oral cancer by conducting a melting curve analysis in a quantitative real-time polymerase chain reaction (qPCR). The assay was performed by a qPCR of DNA from the gargle fluid of 20 patients with oral cancer and 20 healthy control subjects in the training set. The use of these data for the genes BRCA2, CDH13, and MLH1 provided good diagnostic performance. We also applied the assay to a test set of gargle fluid from 10 patients with oral cancer and 10 control subjects, and the results demonstrated the reproducibility of the diagnostic performance. In light of these findings, we propose that this new method could become a noninvasive and convenient screening tool for the detection of oral cancer. |
| format | Article |
| id | doaj-art-3cfc4aba55aa4cd6bc9715d64634dea8 |
| institution | Kabale University |
| issn | 2405-8440 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Heliyon |
| spelling | doaj-art-3cfc4aba55aa4cd6bc9715d64634dea82025-01-30T05:14:42ZengElsevierHeliyon2405-84402025-02-01113e42286Melting curve analyses in the quantitative real-time polymerase chain reaction of methylated/non-methylated DNA toward the detection of oral cancer using gargle fluidYusaku Noma0Hayate Sawayama1Kouta Yamashiro2Keitaro Nishi3Haruka Yoshinaga4Ryota Takaoka5Tsuyoshi Sugiura6Tomonori Sasahira7Tomofumi Hamada8Yasuo Suda9Tatsuo Okui10Department of Maxillofacial Diagnostic and Surgical Science, Field of Oral and Maxillofacial Rehabilitation, Graduate School of Medical and Dental Science, Kagoshima University, Kagoshima, 890-8544, JapanSUDx-Biotec Corporation, C/o Kagoshima University VBL, Kagoshima, 890-0065, JapanDepartment of Maxillofacial Diagnostic and Surgical Science, Field of Oral and Maxillofacial Rehabilitation, Graduate School of Medical and Dental Science, Kagoshima University, Kagoshima, 890-8544, JapanDepartment of Maxillofacial Diagnostic and Surgical Science, Field of Oral and Maxillofacial Rehabilitation, Graduate School of Medical and Dental Science, Kagoshima University, Kagoshima, 890-8544, JapanDepartment of Maxillofacial Diagnostic and Surgical Science, Field of Oral and Maxillofacial Rehabilitation, Graduate School of Medical and Dental Science, Kagoshima University, Kagoshima, 890-8544, JapanDepartment of Maxillofacial Diagnostic and Surgical Science, Field of Oral and Maxillofacial Rehabilitation, Graduate School of Medical and Dental Science, Kagoshima University, Kagoshima, 890-8544, JapanDepartment of Maxillofacial Diagnostic and Surgical Science, Field of Oral and Maxillofacial Rehabilitation, Graduate School of Medical and Dental Science, Kagoshima University, Kagoshima, 890-8544, Japan; Division of Oral and Maxillofacial Oncology and Surgical Sciences, Department of Disease Management Dentistry, Tohoku University Graduate School of Dentistry, Sendai, 980-8575, JapanDepartment of Molecular Oral Pathology and Oncology, Graduate School of Medical and Dental Science, Kagoshima University, Kagoshima, 890-8544, JapanDepartment of Oral & Maxillofacial Surgery, Hakuaikai Medical Cooperation, Sagara Hospital, Kagoshima, 890-8544, JapanSUDx-Biotec Corporation, C/o Kagoshima University VBL, Kagoshima, 890-0065, JapanDepartment of Maxillofacial Diagnostic and Surgical Science, Field of Oral and Maxillofacial Rehabilitation, Graduate School of Medical and Dental Science, Kagoshima University, Kagoshima, 890-8544, Japan; Corresponding author. Department of Maxillofacial Diagnostic and Surgical Science, Field of Oral and Maxillofacial Rehabilitation, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima, 890-8544, Japan.The early detection and early treatment of oral cancer are of paramount importance for improving the patients' prognoses. Our group previously reported a noninvasive test for oral cancer that uses a commercially available methylation-specific multiple ligation-dependent probe amplification (MS-MLPA) method, focusing on the DNA methylation of cells in gargle fluid. However, the conventional MS-MLPA method is complicated, costly, and thus difficult to use for screening and other health examinations. We therefore sought to develop a simple screening method for the diagnosis of oral cancer by conducting a melting curve analysis in a quantitative real-time polymerase chain reaction (qPCR). The assay was performed by a qPCR of DNA from the gargle fluid of 20 patients with oral cancer and 20 healthy control subjects in the training set. The use of these data for the genes BRCA2, CDH13, and MLH1 provided good diagnostic performance. We also applied the assay to a test set of gargle fluid from 10 patients with oral cancer and 10 control subjects, and the results demonstrated the reproducibility of the diagnostic performance. In light of these findings, we propose that this new method could become a noninvasive and convenient screening tool for the detection of oral cancer.http://www.sciencedirect.com/science/article/pii/S2405844025006668Quantitative real-time PCRMelting curve analysisGargle fluidPredictionOral cancerOral squamous cell carcinoma |
| spellingShingle | Yusaku Noma Hayate Sawayama Kouta Yamashiro Keitaro Nishi Haruka Yoshinaga Ryota Takaoka Tsuyoshi Sugiura Tomonori Sasahira Tomofumi Hamada Yasuo Suda Tatsuo Okui Melting curve analyses in the quantitative real-time polymerase chain reaction of methylated/non-methylated DNA toward the detection of oral cancer using gargle fluid Heliyon Quantitative real-time PCR Melting curve analysis Gargle fluid Prediction Oral cancer Oral squamous cell carcinoma |
| title | Melting curve analyses in the quantitative real-time polymerase chain reaction of methylated/non-methylated DNA toward the detection of oral cancer using gargle fluid |
| title_full | Melting curve analyses in the quantitative real-time polymerase chain reaction of methylated/non-methylated DNA toward the detection of oral cancer using gargle fluid |
| title_fullStr | Melting curve analyses in the quantitative real-time polymerase chain reaction of methylated/non-methylated DNA toward the detection of oral cancer using gargle fluid |
| title_full_unstemmed | Melting curve analyses in the quantitative real-time polymerase chain reaction of methylated/non-methylated DNA toward the detection of oral cancer using gargle fluid |
| title_short | Melting curve analyses in the quantitative real-time polymerase chain reaction of methylated/non-methylated DNA toward the detection of oral cancer using gargle fluid |
| title_sort | melting curve analyses in the quantitative real time polymerase chain reaction of methylated non methylated dna toward the detection of oral cancer using gargle fluid |
| topic | Quantitative real-time PCR Melting curve analysis Gargle fluid Prediction Oral cancer Oral squamous cell carcinoma |
| url | http://www.sciencedirect.com/science/article/pii/S2405844025006668 |
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