caa a22 4500 605491291 CHVBK 20210128100509.0 cr unu---uuuuu 210128e20150801xx s 000 0 eng 10.1007/s10147-014-0780-5 doi (NATIONALLICENCE)springer-10.1007/s10147-014-0780-5 A methylation-specific dot blot assay for improving specificity and sensitivity of methylation-specific PCR on DNA methylation analysis [Elektronische Daten] [Vo Lan, Nguyen Trang, Doan Van, Ta Thuan, Ta Van To, Vuong Linh, Nguyen Uyen] Background and objectives: Developing a methylation-specific dot blot assay (MSP-DB) to increase the sensitivity and specificity of simultaneous methylation analysis of multiple genes is the goal of the present study to evaluate the methylation status of GSTP1 and RASSF1A from prostate cancer in Vietnamese males. Methods: The methylation of GSTP1 and RASSF1A was investigated by using the MSP in 50 prostate cancer and 17 benign prostate hyperplasia specimens. The MSP-DB assay that uses a single or multiple probes specifically detected the methylation status of a particular gene or of the two genes GSTP1 and RASSF1A at the same time in a series of samples. The sensitivity and specificity of the MSP-DB were compared to those of the MSP. Results: The probes specifically hybridized with the methylated targets only in the MSP-DB, which allowed detecting GSTP1 and RASSF1A methylation in 23 of 50 and 14 of 50 patients with prostate cancer and in 2 of 17 and 4 of 17 patients with benign prostate hyperplasia. MSP-DB following the MSP assay improved the sensitivity of detection to more than 0.01% methylated status of a given high CpG-rich region. One methylated MSP product corresponding to the GSTP1, lack of methylated cytosine, was clearly detected on gel electrophoresis but barely visible on MSP-DB. Thus, the MSP-DB is suitable to eliminate the risk of false-positive results. Conclusion: The MSP-DB dispels the weakness of MSP and increases the sensitivity to simultaneous methylation analysis of multiple genes. The MSP-DB is advantageous for the promotion of DNA methylation markers in progressing quickly toward clinical application. Japan Society of Clinical Oncology, 2015 DNA methylation nationallicence Glutathione S-transferase P1 (GSTP1) nationallicence RAS - association domain family member 1 (RASSF1A) nationallicence Methylation-specific polymerase chain reaction (MSP) nationallicence Methylation-specific dot blot assay (MSP-DB) nationallicence Lan Vo Faculty of Biology, VNU University of Science, 334, Nguyen Trai, Thanh Xuan, Hanoi, Vietnam aut Trang Nguyen Faculty of Biology, VNU University of Science, 334, Nguyen Trai, Thanh Xuan, Hanoi, Vietnam aut Van Doan Faculty of Biology, VNU University of Science, 334, Nguyen Trai, Thanh Xuan, Hanoi, Vietnam aut Thuan Ta Faculty of Biology, VNU University of Science, 334, Nguyen Trai, Thanh Xuan, Hanoi, Vietnam aut Van To Ta Department of Cytology and Pathology, National Cancer Hospital K, 43 Quan Su, Hoan Kiem, Hanoi, Vietnam aut Linh Vuong Department of Cytology and Pathology, National Cancer Hospital K, 43 Quan Su, Hoan Kiem, Hanoi, Vietnam aut Uyen Nguyen Institute of Microbiology and Biotechnology, Vietnam National University Hanoi, 144, Xuan Thuy, Cau Giay, Hanoi, Vietnam aut International Journal of Clinical Oncology Springer Japan 20/4(2015-08-01), 839-845 1341-9625 20:4<839 2015 20 10147 https://doi.org/10.1007/s10147-014-0780-5 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10147-014-0780-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Lan Vo Faculty of Biology, VNU University of Science, 334, Nguyen Trai, Thanh Xuan, Hanoi, Vietnam aut NATIONALLICENCE 700 1- Trang Nguyen Faculty of Biology, VNU University of Science, 334, Nguyen Trai, Thanh Xuan, Hanoi, Vietnam aut NATIONALLICENCE 700 1- Van Doan Faculty of Biology, VNU University of Science, 334, Nguyen Trai, Thanh Xuan, Hanoi, Vietnam aut NATIONALLICENCE 700 1- Thuan Ta Faculty of Biology, VNU University of Science, 334, Nguyen Trai, Thanh Xuan, Hanoi, Vietnam aut NATIONALLICENCE 700 1- Van To Ta Department of Cytology and Pathology, National Cancer Hospital K, 43 Quan Su, Hoan Kiem, Hanoi, Vietnam aut NATIONALLICENCE 700 1- Linh Vuong Department of Cytology and Pathology, National Cancer Hospital K, 43 Quan Su, Hoan Kiem, Hanoi, Vietnam aut NATIONALLICENCE 700 1- Uyen Nguyen Institute of Microbiology and Biotechnology, Vietnam National University Hanoi, 144, Xuan Thuy, Cau Giay, Hanoi, Vietnam aut NATIONALLICENCE 773 0- International Journal of Clinical Oncology Springer Japan 20/4(2015-08-01), 839-845 1341-9625 20:4<839 2015 20 10147