MS-HRM to Detect Serum DNA Methylation of Intrauterine Growth Retardation Children

Du, Yongmei; Zhou, Youxia; Wu, Qian

doi:10.4236/eng.2012.410b027

Cited by 2 publications

(3 citation statements)

References 17 publications

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“…In the present MS-HRM assay, melting temperature was obtained as a T m value in the T m calling analysis. This refers to the temperature at which half of the double-stranded DNA melts into single strands, thus undergoing a sharp decline in fluorescence intensity ( 8 , 9 ). Fig.…”

Section: Resultsmentioning

confidence: 99%

“…At present, there are various methylation detection methods that are in use, including methylation-specific polymerase chain reaction ( 6 ), methylight ( 7 ), methylation-sensitive high-resolution melting (MS-HRM) ( 8 ), pyrosequencing ( 9 ), and microarray technologies ( 10 ). Known as a sensitive and more efficient technology for detecting single nucleotide variations ( 11 ), MS-HRM is considered to be a novel technology that would provide clinically-relevant sensitivity and rapidity for gene methylation screening.…”

Section: Introductionmentioning

confidence: 99%

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Quantitative assessment of gene promoter methylation in non‑small cell lung cancer using methylation‑sensitive high‑resolution melting

Liu

Zhang

et al. 2018

Oncol Lett

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DNA methylation is closely associated with aberrant epigenetic changes. Previous studies have identified various genes associated with non-small cell lung cancer (NSCLC), but the precise combination responsible for its etiology is still debated. The aim of the present study was to select a new set of NSCLC-related genes using methylation-sensitive high-resolution melting. The promoter methylation status of six selected genes, consisting of protocadherin γ subfamily B, 6 (PCDHGB6), homeobox A9 (HOXA9), O6-methylguanine-DNA methyltransferase (MGMT), microRNA (miR)-126, suppressor of cytokine signaling 3 (SOCS3) and Ras association domain family member 5, also termed NORE1A, was evaluated in 54 NSCLC patients. From these samples, genome-wide DNA was extracted and bisulfite conversion was performed along with fluorogenic quantitative polymerase chain reaction to detect methylation values of the six selected promoters. The present results revealed frequent methylation on PCDHGB6, HOXA9 and miR-126, which contrasted with infrequent methylation on MGMT. The results indicated no methylation on either SOCS3 or NORE1A. The sensitivity and specificity of the methylation assessment were 85.2 and 81.5%, respectively, and the analysis results were validated by pyrosequencing. Furthermore, minute comparison of the association between DNA methylation and clinical features was performed. Overall, these results may provide potential information for the development of better clinical diagnostics and more targeted and effective therapies for NSCLC.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Quantitative assessment of gene promoter methylation in non‑small cell lung cancer using methylation‑sensitive high‑resolution melting

Liu

Zhang

et al. 2018

Oncol Lett

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“…There are some reports applying MS-HRM for methylation detection in clinical samples. For example, the detection of serum DNA methylation in intrauterine growth retardation infants [58], in pregnant women [59], and in nasopharyngeal carcinoma patients [60]. Moreover, HRM is suitable in many clinical applications such as point mutation detection, single-nucleotide polymorphism (SNP), and microbial genotyping [61].…”

Section: Discussionmentioning

confidence: 99%

Serum cell-free DNA methylation of OPCML and HOXD9 as a biomarker that may aid in differential diagnosis between cholangiocarcinoma and other biliary diseases

et al. 2019

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Background Cholangiocarcinoma (CCA) is a fatal cancer of the bile duct epithelial cell lining. The misdiagnosis of CCA and other biliary diseases may occur due to the similarity of clinical manifestations and blood tests resulting in inappropriate or delayed treatment. Thus, an accurate and less-invasive method for differentiating CCA from other biliary diseases is inevitable. Methods We quantified methylation of OPCML , HOXA9 , and HOXD9 in serum cell-free DNA (cfDNA) of CCA patients and other biliary diseases using methylation-sensitive high-resolution melting (MS-HRM). Their potency as differential biomarkers between CCA and other biliary diseases was also evaluated by using receiver operating characteristic (ROC) curves. Results The significant difference of methylation levels of OPCML and HOXD9 was observed in serum cfDNA of CCA compared to other biliary diseases. Assessment of serum cfDNA methylation of OPCML and HOXD9 as differential biomarkers of CCA and other biliary diseases showed the area under curve (AUC) of 0.850 (0.759–0.941) for OPCML which sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy were 80.00%, 90.00%, 88.88%, 81.81%, and 85.00%, respectively. The AUC of HOXD9 was 0.789 (0.686–0.892) with sensitivity, specificity, PPV, NPV, and accuracy of 67.50%, 90.00%, 87.09%, 73.46%, and 78.75%, respectively. The combined marker between OPCML and HOXD9 showed sensitivity, specificity, PPV, and NPV of 62.50%, 100%, 100%, and 72.72%, respectively, which may be helpful to prevent a misdiagnosis between CCA and other biliary diseases. Conclusions Our findings suggest the application of serum cfDNA methylation of OPCML and HOXD9 for differential diagnosis of CCA and other biliary diseases due to its less invasiveness and clinically practical method which may benefit the patients by preventing the misdiagnosis of CCA and avoiding unnecessary surgical intervention. Electronic supplementary material The online version of this article (10.1186/s13148-019-0634-0) contains supplementary material, which is available to authorized users.

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MS-HRM to Detect Serum DNA Methylation of Intrauterine Growth Retardation Children

Cited by 2 publications

References 17 publications

Quantitative assessment of gene promoter methylation in non‑small cell lung cancer using methylation‑sensitive high‑resolution melting

Quantitative assessment of gene promoter methylation in non‑small cell lung cancer using methylation‑sensitive high‑resolution melting

Serum cell-free DNA methylation of OPCML and HOXD9 as a biomarker that may aid in differential diagnosis between cholangiocarcinoma and other biliary diseases

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