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. 2021 Mar 12;2021(3):CD013316. doi: 10.1002/14651858.CD013316.pub2
Study Technique Sample type CpGs analysed (PCR‐based tests) Threshold for methylated HR (95% confidence interval) Data source
Almuqate 2018 MS‐RE‐qPCR NR NR > 5% 4.17 (1.78 to 9.75) Adjusted HR (for age, sex and surgery)
MS‐RE‐qPCR NR NR > 9% 3.57 (1.67 to 7.62) Adjusted HR (for age, sex and surgery)
Bady 2012 (E‐GBM) Bead array Frozen 31 and 83 > 0.358 3.28 (1.68 to 6.41) Unadjusted HR
Bead array Frozen 78–84 > 10% 5.56 (1.25 to 25.0) Unadjusted HR
PSQ Frozen 74–78 > 7.28% 2.20 (1.12 to 4.31) Unadjusted HR
Bady 2012 (M‐GBM) Bead array Frozen 31 and 83 > 0.358 6.46 (2.41 to 17.3) IPD
MSP NR 76–80 and 84–86 NR 7.21 (2.37 to 22.0) IPD
Barault 2015 Methyl‐beaming FFPE 79–83 > 40.2% 2.78 (1.85 to 5.26) Unadjusted HR
PSQ FFPE 76–81 > 29.6% 2.63 (1.43 to 4.55) Unadjusted HR
Barbagallo 2014 MSP FFPE 76–80 and 84–87 Including weakly 3.68 (1.66 to 8.18) KM curves (Fig 3A or 3C, Barbagallo 2014)
MSP FFPE 76–80 and 84–87 Excluding weakly 1.90 (0.72 to 4.99) KM curves (Fig 3C, Barbagallo 2014)
PSQ FFPE NR > 9% 3.73 (1.68 to 8.28) KM curves (Fig 3A or 3B, Barbagallo 2014)
PSQ FFPE NR > 25% 1.99 (0.92 to 4.32) KM curves (Fig 3B, Barbagallo 2014)
Bell 2017 QF‐IHC (AQUA) FFPE N/A > Median 1.84 (1.38 to 2.43) Adjusted HR (for age, KPS, resection status and treatment)
qMSP NR NR > 8 1.77 (1.28 to 2.44) Adjusted HR (for age, KPS, resection status and treatment)
Brigliadori 2016 PSQ FFPE 74–83 > 9% 1.92 (1.17 to 3.14) KM curves (Fig 1; stratified by extent of resection, Brigliadori 2016)
PSQ FFPE 74–83 > 29% 3.02 (1.72 to 5.29) KM curves (Fig 2; stratified by extent of resection, Brigliadori 2016)
Chai 2018 (7‐site cohort) PSQ Frozen 72–78 > 12% 2.94 (1.12 to 7.69) Unadjusted HR
PSQ Frozen 74–78 > 12% 2.94 (1.12 to 7.69) Unadjusted HR
PSQ Frozen 75–78 > 12% 2.94 (1.12 to 7.69) Unadjusted HR
Chai 2018 (8‐site cohort) PSQ Frozen 75–78 > 13% 2.70 (1.37 to 5.26) Unadjusted HR
PSQ Frozen 75–82 > 12% 3.03 (1.54 to 6.25) Unadjusted HR
PSQ Frozen 76–79 > 11% 2.13 (1.09 to 4.17) Unadjusted HR
Dahlrot 2018 (NS cohort) DIF FFPE N/A < 0.2 1.60 (0.95 to 2.71) Unadjusted HR
PSQ FFPE 74–78 > 9% 1.42 (0.84 to 2.40) Unadjusted HR
Dahlrot 2018 (RSD cohort) DIF FFPE N/A < 0.2 2.00 (1.32 to 3.02) Unadjusted HR
PSQ FFPE 74–78 > 10% 1.58 (1.14 to 2.19) KM curves (Fig 3A, Dahlrot 2018)
Dunn 2009 PSQ Frozen, smear, FFPE or a combination 72–83 > 9% 3.57 (2.24 to 5.70) KM curves (Fig 2B, Dunn 2009)
PSQ Frozen, smear, FFPE or a combination 72–83 > 20% 4.25 (2.57 to 7.05) KM curves (Fig 2D, Dunn 2009)
PSQ Frozen, smear, FFPE or a combination 72–83 > 29% 4.03 (2.30 to 7.07) KM curves (Fig 2F, Dunn 2009)
PSQ Frozen, smear, FFPE or a combination 72–83 > 35% 3.64 (1.99 to 6.67) KM curves (Fig 2D, Dunn 2009)
PSQ Frozen, smear, FFPE or a combination 72–83 Cluster 1 vs 2 and 3 4.44 (2.58 to 7.66) KM curves (Fig Suppl 4C, Dunn 2009)
PSQ Frozen, smear, FFPE or a combination 72–83 Cluster 1 and 2 vs 3 3.59 (2.26 to 5.69) KM curves (Fig Suppl 4C, Dunn 2009)
Felsberg 2009 IHC FFPE N/A < 10% 1.26 (0.70 to 2.25) KM curves (Fig Suppl 1C, 2nd column, Felsberg 2009)
MSP Frozen (14 FFPE) NR NR 2.23 (1.29 to 3.84) KM curves (Fig Suppl 1A, 2nd column, Felsberg 2009)
PCR‐mRNA Frozen (14 FFPE) N/A < 50% 2.66 (0.94 to 7.53) KM curves (Fig Suppl 1B, 2nd column, Felsberg 2009)
Havik 2012 MSP Frozen 76–80 and 84–87 NR 2.02 (1.08 to 3.77) KM curves (Fig 2A, Johannessen 2018)
PCR‐HRM Frozen 72–83 NR 1.32 (0.64 to 2.68) KM curves (Fig 2B, Johannessen 2018)
PSQ Frozen 74–78 > 2.68% 1.85 (1.02 to 3.35) KM curves (Fig 1B, Havik 2012)
PSQ Frozen 74–78 > 6% 2.30 (1.21 to 4.38) Unadjusted HR
PSQ Frozen 74–78 > 7% 2.33 (1.19 to 4.57) Unadjusted HR
PSQ Frozen 74–78 > 8% 2.33 (1.19 to 4.57) Unadjusted HR
PSQ Frozen 74–78 > 9% 2.30 (0.99 to 5.33) Unadjusted HR
PSQ Frozen 76–79 > 6% 2.30 (1.21 to 4.38) Unadjusted HR
PSQ Frozen 76–79 > 7% 2.33 (1.19 to 4.57) Unadjusted HR
PSQ Frozen 76–79 > 8% 1.90 (0.99 to 3.65) Unadjusted HR
PSQ Frozen 76–79 > 9% 1.90 (0.99 to 3.65) Unadjusted HR
qMSP Frozen 71–73 and 75–86 NR 1.72 (0.91 to 3.22) KM curves (Fig 2C, Johannessen 2018)
qMSP Frozen 71–86 > 0% 1.66 (0.97 to 2.83) KM curves (Fig 1A, Havik 2012)
Hsu 2015 IHC FFPE N/A < 10% 2.12 (1.32 to 3.42) Unadjusted HR
MSP FFPE 76–80 and 84–87 NR 2.39 (1.42 to 4.02) Unadjusted HR
PSQ FFPE 76–79 > 5% 2.66 (1.49 to 4.76) Unadjusted HR
qMSP FFPE 77–80 and 84–87 > 0.04% 2.75 (1.51 to 5.04) Unadjusted HR
qMSP FFPE 77–80 and 84–87 > 0.1% 2.83 (1.85 to 4.33) IPD
Karayan‐Tapon 2010 IHC FFPE N/A < 15.5% 1.26 (0.77 to 2.06) KM curves (Fig 3B, Karayan‐Tapon 2010)
MSP Frozen 76–80 and 84–87 NR 2.32 (1.39 to 3.87) KM curves (Fig 1A, Karayan‐Tapon 2010)
PCR‐mRNA Frozen N/A < 0.39 1.68 (1.04 to 2.73) KM curves (Fig 3A, Karayan‐Tapon 2010)
PSQ Frozen 74 > 5.5% 3.26 (1.91 to 5.59) KM curves (Fig Elec Suppl 1B, Karayan‐Tapon 2010)
PSQ Frozen 74–78 > 8.0% 3.35 (1.95 to 5.73) KM curves (Fig 2B; cpg mean, Karayan‐Tapon 2010)
PSQ Frozen 75 > 8.7% 3.26 (1.91 to 5.57) KM curves (Fig Elec Suppl 1B, Karayan‐Tapon 2010)
PSQ Frozen 76 > 8.0% 2.78 (1.64 to 4.74) KM curves (Fig Elec Suppl 1B, Karayan‐Tapon 2010)
PSQ Frozen 77 > 7.85% 3.65 (2.10 to 6.34) KM curves (Fig 2B, Karayan‐Tapon 2010)
PSQ Frozen 78 > 7.8% 2.70 (1.59 to 4.58) KM curves (Fig Elec Suppl 1B, Karayan‐Tapon 2010)
SQ‐MSP Frozen 76–80 and 84–87 > 35 2.75 (1.66 to 4.53) KM curves (Fig 1B, Karayan‐Tapon 2010)
Kim 2016 MSP FFPE 76–80 and 84–87 NR 7.66 (2.82 to 20.8) KM curves (Fig 2G, Kim 2016)
PSQ FFPE 74–78 > 9% 7.66 (2.82 to 20.8) KM curves (Fig 2G, Kim 2016)
Kristensen 2016 IHC FFPE N/A At 0% 1.58 (1.12 to 2.22) Unadjusted HR
PSQ Frozen NR > 10% 1.80 (1.23 to 2.62) Unadjusted HR
qMSP‐PSQ Frozen NR > 0.1% 1.64 (1.15 to 2.33) Unadjusted HR
qMSP‐PSQ Frozen NR > 5% 1.66 (1.02 to 2.71) KM curves (Fig 5a, Kristensen 2016)
qMSP‐PSQ Frozen NR > 20% 1.52 (0.77 to 3.00) KM curves (Fig 5a, Kristensen 2016)
Lalezari 2013 IHC FFPE N/A < 30% 1.74 (1.39 to 2.16) KM curves (Fig 1A, Lalezari 2013)
MSP FFPE 76–80 and 84–87 NR 2.13 (1.67 to 2.78) Adjusted HR (for age, sex, KPS, extent of resection, bevacizumab treatment at any time and IDH1R132 mutation status)
PSQ FFPE 72–95 NR 2.06 (1.62 to 2.62) KM curves (Fig 1E; unadjusted, Lalezari 2013)
Lattanzio 2015 MSP FFPE 76–80 and 84–87 NR 1.45 (0.76 to 2.76) KM curves (Fig 3B, Lattanzio 2015)
MSP Frozen 76–80 and 84–87 NR 2.27 (1.21 to 4.26) KM curves (Fig 3A, Lattanzio 2015)
PSQ FFPE 72–80 ≥ 9% 2.09 (1.09 to 3.99) KM curves (Fig 3D, Lattanzio 2015)
PSQ Frozen 72–80 ≥ 9% 2.25 (1.19 to 4.25) KM curves (Fig 3C, Lattanzio 2015)
Lechapt‐Zalcman 2012 IHC FFPE N/A < 15% 1.99 (1.15 to 3.42) KM curves (Fig 4d; unadjusted, Lechapt‐Zalcman 2012)
MSP FFPE 76–80 and 84–87 NR 1.78 (1.03 to 3.09) KM curves (Fig 4c; unadjusted, Lechapt‐Zalcman 2012)
McDonald 2013 MSP FFPE 76–80 NR 1.64 (0.95 to 2.83) IPD
PSQ FFPE 74–78 > 8% 1.96 (1.16 to 3.33) Unadjusted HR
Melguizo 2012 IHC FFPE N/A < 25% 1.11 (0.69 to 1.77) KM curves (Fig 4B, Melguizo 2012)
MSP NR 76–80 and 84–87 NR 1.77 (1.06 to 2.95) KM curves (Fig 4A, Melguizo 2012)
Nguyen 2015 FSQ‐MS‐PCR Frozen or FFPE 76–80 and 84–87 > 15% 2.68 (1.70 to 4.21) KM curves (Fig 3D, Nguyen 2015)
FSQ‐MS‐PCR Frozen or FFPE 76–80 and 84–87 > 60% 2.25 (1.31 to 3.87) KM curves (Fig 3D, Nguyen 2015)
Park 2011 MS‐MLPA 50% frozen and 50% FFPE NR > 0.1% 2.38 (1.11 to 5.10) KM curves (Fig 2B, Park 2011)
MS‐MLPA 50% frozen and 50% FFPE NR > 0.2 1.88 (0.86 to 4.11) KM curves (Fig 2B, Park 2011)
MSP 50% frozen and 50% FFPE 76–80 and 84–86 NR 4.53 (1.58 to 12.9) KM curves (Fig 2A, Park 2011)
Quillien 2014 (test) IHC FFPE N/A < 23% 2.33 (1.44 to 3.74) Adjusted HR (for age and KPS)
MSP Frozen 76–80 and 84–87 NR 2.70 (1.65 to 4.43) Adjusted HR (for age and KPS)
MethyLight‐MSP Frozen 75–86 > 0 1.67 (1.00 to 2.77) Adjusted HR (for age and KPS)
PCR‐HRM Frozen 70–83 > 50% 1.92 (1.12 to 3.29) Adjusted HR (for age and KPS)
PSQ Frozen 74 > 4% 2.44 (1.51 to 3.95) Adjusted HR (for age and KPS)
PSQ Frozen 74 > 8% 2.22 (1.30 to 3.79) Adjusted HR (for age and KPS)
PSQ Frozen 74–78 > 8% 3.13 (1.86 to 5.25) Adjusted HR (for age and KPS)
PSQ Frozen 74–78 > 9% 3.13 (1.81 to 5.38) Adjusted HR (for age and KPS)
PSQ Frozen 74–89 > 11% 3.13 (1.84 to 5.30) Adjusted HR (for age and KPS)
PSQ Frozen 75 > 11% 3.23 (1.87 to 5.55) Adjusted HR (for age and KPS)
PSQ Frozen 75–79 > 8% 2.94 (1.71 to 5.06) Adjusted HR (for age and KPS)
PSQ Frozen 76 > 4% 2.63 (1.61 to 4.31) Adjusted HR (for age and KPS)
PSQ Frozen 76 > 5% 2.78 (1.62 to 4.75) Adjusted HR (for age and KPS)
PSQ Frozen 76–79 > 8% 2.86 (1.66 to 4.92) Adjusted HR (for age and KPS)
PSQ Frozen 76–80 > 9% 3.03 (1.74 to 5.26) Adjusted HR (for age and KPS)
PSQ Frozen 77 > 6% 3.13 (1.87 to 5.23) Adjusted HR (for age and KPS)
PSQ Frozen 77 > 7% 2.78 (1.63 to 4.74) Adjusted HR (for age and KPS)
PSQ Frozen 77–81 > 8% 2.70 (1.59 to 4.59) Adjusted HR (for age and KPS)
PSQ Frozen 78 > 4% 2.33 (1.38 to 3.92) Adjusted HR (for age and KPS)
PSQ Frozen 78 > 5% 2.50 (1.51 to 4.13) Adjusted HR (for age and KPS)
PSQ Frozen 78–82 > 9% 2.86 (1.68 to 4.86) Adjusted HR (for age and KPS)
PSQ Frozen 79 > 7% 2.78 (1.61 to 4.80) Adjusted HR (for age and KPS)
PSQ Frozen 79–83 > 8% 2.86 (1.68 to 4.86) Adjusted HR (for age and KPS)
PSQ Frozen 80 > 4% 2.56 (1.54 to 4.28) Adjusted HR (for age and KPS)
PSQ Frozen 81 > 8% 2.44 (1.44 to 4.14) Adjusted HR (for age and KPS)
PSQ Frozen 82 > 16% 2.94 (1.70 to 5.08) Adjusted HR (for age and KPS)
PSQ Frozen 83 > 10% 2.78 (1.65 to 4.66) Adjusted HR (for age and KPS)
PSQ Frozen 84 > 9% 3.23 (1.86 to 5.59) Adjusted HR (for age and KPS)
PSQ Frozen 84–88 > 17% 3.23 (1.86 to 5.58) Adjusted HR (for age and KPS)
PSQ Frozen 84–89 > 22% 3.23 (1.85 to 5.62) Adjusted HR (for age and KPS)
PSQ Frozen 85 > 5% 2.50 (1.52 to 4.11) Adjusted HR (for age and KPS)
PSQ Frozen 85–89 > 13% 2.94 (1.76 to 4.92) Adjusted HR (for age and KPS)
PSQ Frozen 86 > 11% 2.78 (1.65 to 4.69) Adjusted HR (for age and KPS)
PSQ Frozen 87 > 25% 3.03 (1.75 to 5.24) Adjusted HR (for age and KPS)
PSQ Frozen 88 > 4% 2.27 (1.38 to 3.75) Adjusted HR (for age and KPS)
PSQ Frozen 89 > 12% 3.23 (1.91 to 5.46) Adjusted HR (for age and KPS)
Quillien 2014 (validation) PSQ FFPE 74–78 > 9% 3.70 (1.71 to 8.01) Unadjusted HR
PSQ FFPE 74–78 > 10% 2.86 (1.42 to 5.74) Unadjusted HR
PSQ FFPE 74–78 > 28% 2.27 (0.98 to 5.29) Unadjusted HR
Quillien 2016 PSQ FFPE 74–78 > 6% 3.23 (2.02 to 5.16) Adjusted HR (for age and KPS)
PSQ FFPE 74–78 > 8% 4.00 (2.30 to 6.97) Adjusted HR (for age and KPS)
PSQ FFPE 74–78 > 12% 4.17 (2.35 to 7.38) Adjusted HR (for age and KPS)
PSQ FFPE 74–78 > 13% 4.35 (2.41 to 7.83) Adjusted HR (for age and KPS)
PSQ FFPE 74–78 > 16% 4.55 (2.48 to 8.34) Adjusted HR (for age and KPS)
PSQ Frozen 74–78 > 6% 4.00 (2.30 to 6.97) Adjusted HR (for age and KPS)
PSQ Frozen 74–78 > 8% 3.57 (2.14 to 5.95) Adjusted HR (for age and KPS)
PSQ Frozen 74–78 > 12% or 13% 3.45 (2.10 to 5.66) Adjusted HR (for age and KPS)
PSQ Frozen 74–78 > 12% 3.70 (2.19 to 6.26) Adjusted HR (for age and KPS)
PSQ Frozen 74–78 > 16% 3.13 (1.94 to 5.04) Adjusted HR (for age and KPS)
PSQ Frozen 76–79 > 8% 3.33 (2.06 to 5.40) Adjusted HR (for age and KPS)
PSQ Frozen 76–79 > 12% 3.33 (2.06 to 5.40) Adjusted HR (for age and KPS)
SQ‐MSP FFPE 76–80 and 84–87 > 12% 3.33 (2.06 to 5.40) Adjusted HR (for age and KPS)
SQ‐MSP FFPE 76–80 and 84–87 > 13% 3.33 (2.06 to 5.40) Adjusted HR (for age and KPS)
SQ‐MSP FFPE 76–80 and 84–87 > 23% 4.17 (2.35 to 7.38) Adjusted HR (for age and KPS)
SQ‐MSP Frozen 76–80 and 84–87 > 13% 2.86 (1.79 to 4.57) Adjusted HR (for age and KPS)
SQ‐MSP Frozen 76–80 and 84–87 > 23% 2.17 (1.38 to 3.44) Adjusted HR (for age and KPS)
Thon 2017 MSP Frozen 76–80 and 84–87 NR 3.33 (1.82 to 6.25) Unadjusted HR
Sequencing Frozen 75–99 (unclear) > 50% 3.33 (1.82 to 6.25) Unadjusted HR
Yamashita 2018 MSP Frozen 76–80 and 84–87 NR 2.36 (1.62 to 5.05) Unadjusted HR
PCR‐HRM Frozen 72–89 > 5% 2.36 (1.43 to 3.90) KM curves (Fig Suppl 13, Yamashita 2018)
PCR‐HRM Frozen 72–89 > 8% 2.05 (1.24 to 3.39) KM curves (Fig Suppl 13, Yamashita 2018)
PCR‐HRM Frozen 72–89 > 10% 2.51 (1.63 to 4.83) Unadjusted HR
PCR‐HRM Frozen 72–89 > 12% 2.61 (1.54 to 4.42) KM curves (Fig Suppl 13, Yamashita 2018)
PCR‐HRM Frozen 72–89 > 15% 2.39 (1.41 to 4.05) KM curves (Fig Suppl 13, Yamashita 2018)
Yang 2012 IHC FFPE N/A < 10% 1.07 (0.35 to 3.31) IPD
MSP FFPE 76–80 and 84–87 NR 1.35 (0.44 to 4.16) IPD
Yoshioka 2018 SQ‐MSP Frozen 76–80 and 84–87 > 0 2.72 (1.28 to 5.74) KM curves (Fig 2A, Yoshioka 2018)
SQ‐MSP Frozen 76–80 and 84–87 > 2 2.18 (1.20 to 3.97) KM curves (Fig 2B, Yoshioka 2018)
SQ‐MSP Frozen 76–80 and 84–87 > 4 1.85 (1.07 to 3.18) KM curves (Fig 2C, Yoshioka 2018)
SQ‐MSP Frozen 76–80 and 84–87 > 6 1.83 (1.10 to 3.04) KM curves (Fig 2D, Yoshioka 2018)
SQ‐MSP Frozen 76–80 and 84–87 > 8 1.71 (1.00 to 2.93) KM curves (Fig 2E, Yoshioka 2018)
CpG: 5'‐cytosine‐phosphate‐guanine‐3'; DIF: double immunofluorescence; FFPE: formalin‐fixed paraffin‐embedded; FSQ‐MS‐PCR: fluorescent semi‐quantitative methylation‐specific polymerase chain reaction; HR: hazard ratio; IDH: isocitrate dehydrogenase; IHC: immunohistochemistry; IPD: individual participant data; KM: Kaplan‐Meier; KPS: Karnofsky performance status; MS‐MLPA: methylation‐specific multiplex ligation‐dependent probe amplification; MS‐RE‐qPCR: methylation‐specific restriction enzyme quantitative polymerase chain reaction; MSP: methylation‐specific polymerase chain reaction; N/A: not applicable; NR: not reported; PCR: polymerase chain reaction; PCR‐HRM: polymerase chain reaction with high‐resolution melting; PCR‐mRNA: polymerase chain reaction‐messenger ribonucleic acid; PSQ: pyrosequencing; QF‐IHC: quantitative fluorescence immunohistochemistry; qMSP: quantitative methylation‐specific polymerase chain reaction; qMSP‐PSQ: quantitative methylation‐specific polymerase chain reaction with pyrosequencing; SQ‐MSP: semi‐quantitative methylation‐specific polymerase chain reaction.