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首页> 外文期刊>Cancer: A Journal of the American Cancer Society >Comparative assessment of 5 methods (methylation-specific polymerase chain reaction, methylight, pyrosequencing, methylation-sensitive high-resolution melting, and immunohistochemistry) to analyze O6-methylguanine-DNA- methyltranferase in a series of 100 glioblastoma patients
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Comparative assessment of 5 methods (methylation-specific polymerase chain reaction, methylight, pyrosequencing, methylation-sensitive high-resolution melting, and immunohistochemistry) to analyze O6-methylguanine-DNA- methyltranferase in a series of 100 glioblastoma patients

机译：比较评估5种方法（100例胶质母细胞瘤患者中O6-甲基鸟嘌呤-DNA-甲基转移酶）的5种方法（甲基化特异性聚合酶链反应，甲基维，焦磷酸测序，甲基化敏感的高分辨率熔解和免疫组化）

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Background: There is a strong need to determine the best technique for O 6-methylguanine-DNA-methyltranferase (MGMT) analysis, because MGMT status is currently used in clinical trials and occasionally in routine clinical practice for glioblastoma patients. Methods: The authors compared analytical performances and predictive values of 5 techniques in a series of 100 glioblastoma patients who received standard of care treatment (Stupp protocol). Results: MGMT promoter was considered methylated in 33%, 33%, 42%, and 60% of patients by methylation-sensitive high-resolution melting, MethyLight, pyrosequencing (with an optimal risk cutoff at 8% for the average percentage of the 5 CpGs tested), and methylation-specific polymerase chain reaction (MS-PCR), respectively. Fifty-nine percent of the samples had 23% (the optimal risk cutoff) of MGMT-positive tumor cells. The best predictive values for overall survival (OS), after adjustment for age and performance status, were obtained by pyrosequencing (hazard ratio [HR], 0.32; P .0001), MS-PCR (HR, 0.37; P .0001), and immunohistochemistry (HR, 0.43; P =.0005) as compared with methylation-sensitive high-resolution melting (HR, 0.52 P =.02) and MethyLight (HR, 0.6; P =.05). For progression-free survival (PFS), the best predictive values were obtained with pyrosequencing (HR, 0.35; P .0001), methylation-sensitive high-resolution melting (HR, 0.46; P =.002), and MS-PCR (HR, 0.49; P =.002). Combining pyrosequencing and immunohistochemistry slightly improved predictive power for OS, but not for PFS. Poor reproducibility and interobserver variability were, however, observed for immunohistochemistry. Conclusions: Good prediction of survival in addition to high reproducibility and sensitivity made pyrosequencing the best among the 5 techniques tested in this study.
机译：背景：迫切需要确定用于O 6-甲基鸟嘌呤-DNA-甲基转移酶（MGMT）分析的最佳技术，因为MGMT状态目前已在临床试验中使用，有时还用于胶质母细胞瘤患者的常规临床实践中。方法：作者比较了100例接受标准治疗（Stupp方案）的胶质母细胞瘤患者的5种技术的分析性能和预测价值。结果：通过甲基化敏感的高分辨率融解，MethyLight，焦磷酸测序，MGMT启动子被认为在33％，33％，42％和60％的患者中被甲基化（对于5个平均百分比，最佳风险阈值为8％ CpGs测试）和甲基化特异性聚合酶链反应（MS-PCR）。 59％的样本的MGMT阳性肿瘤细胞＜23％（最佳风险临界值）。通过焦磷酸测序（风险比[HR]，0.32; P <.0001），MS-PCR（HR，0.37; P <.0001），在校正年龄和表现状态后，获得最佳的总生存期（OS）预测值。 ），免疫组化（HR，0.43; P = .0005）与甲基化敏感的高分辨率熔解（HR，0.52 P = .02）和MethyLight（HR，0.6; P = .05）进行比较。对于无进展生存期（PFS），焦磷酸测序（HR，0.35; P <.0001），甲基化敏感的高分辨率熔解（HR，0.46; P = .002）和MS-PCR可获得最佳预测值（HR，0.49; P = .002）。焦磷酸测序和免疫组织化学相结合可稍微改善OS的预测能力，但不能改善PFS。然而，对于免疫组织化学观察到差的再现性和观察者间变异性。结论：除了高重现性和敏感性外，对生存的良好预测使焦磷酸测序成为本研究中测试的5种技术中最好的。

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