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. 2025 Apr 23;23:465. doi: 10.1186/s12967-025-06495-2

Table 2.

Summary of recent liquid biopsy-based DNA methylation studies in breast cancer

Study (authors) Cancer types Markers Sample Detection method (s) No. of cases No. of controls AUC Sens (%) Stage I/II Sens (%) Spec (%)
For detection
Mijnes et al. [31] Breast SPAG6, NKX2-6, ITIH5, PER1 Plasma 450 K array, pyrosequencing 111 14 0.729 63 80
Moss et al. [72] Breast KRT19, LMX1B, ZNF296 Plasma TBS 30 64 0.913 80 97
Liu et al. [64] Breast

10 cfDNA markers

(RYR2, RYR3, GABRB3, DCDC2 C, AC096570.1, LINC00923)

Plasma WGBS 101 102 0.81
Wang et al. [28] Breast CCDC181, GCM2, ITPRIPL1 Plasma EPIC (850 K) array, qMSP 199 247 0.961 92.9 87.5
Zhang et al. [43] Breast 26 cfDNA markers Plasma TBS 204 129 0.945 87.76 87.5 92.73
Gao et al. [13] Breast 15 ctDNA DMRs Plasma WGBS, ddPCR 123 40 0.967 87 100
Kresovich et al. [35] Breast 19 CpGs & 5 DNA methylation estimators Blood 450 K array 1728 1375 0.69
Chung et al. [122] Breast 49 genomic regions Buffy coat RRBS 340 340 0.86
Wang et al. [32] Breast

4 CpG markers

(cg16652347, cg13828440, cg11754974 & cg18637238)

PBMC 850 K (EPIC) array, pyrosequencing, TBS 491 290 0.918 80.3 85.7 89.1
Lee et al. [34] Breast 51 CpG sites Blood EPIC array 256 268 0.827 75 78
Liu et al. [71] Multiple (> 50 cancer types, including BC) A classifier developed based on targeted methylation panel of > 100,000 methylation regions (refer to reference [71]) Plasma TBS 2482 4207 54.9 18 (for all cancer types); < 5 (for BC) 99.3
Klein et al. [40] Multiple (> 50 cancer types, including BC) A refined assay and classifiers based on Liu et al. (refer to reference [40]) Plasma TBS 2823 1254 51.5 16.8 (for all cancer types); 2.6 (for BC) 99.5
Jamshidi et al. [82] Multiple (> 20 cancer types, including BC) Whole-genome methylation classifier (refer to reference [82]) Plasma WGBS 1628 1172 0.63 34 98
Gao et al. [13] Breast 12 ctDNA DMRs Plasma WGBS 78 (ER +) 43 (ER−) 0.780 73 87
Manoochehri et al. [30] Breast 3 DMRs overlapping with SPAG6, LINC10606, TBCD/ZNF750 Plasma 450 K & EPIC arrays, ddPCR 262 (TNBC) 84 0.74

AUC area under the curve, BC breast cancer, ddPCR droplet digital PCR, DMRs differentially methylated regions, ER estrogen receptor, qMSP quantitative methylation-specific PCR, RRBS reduced representation bisulfite sequencing, Sens sensitivity, Spec specificity, TBS targeted bisulfite sequencing, TNBC triple-negative breast cancer, WGBS whole-genome bisulfite sequencing