• 10 patients

• AJCC stage II-IV BTCs patients (4 gallbladder carcinomas and 6 with CCA)

Overall mutation concordance:

• 90% (9/10) concordant mutations between bile cfDNA and tumor tissue DNA

High mutational concordance:

• 70% (7/10) patients exhibited > 50% mutational concordance between bile cfDNA and tumor tissue DNA

Bile-based liquid biopsy features high concordance with blood samples and tumor tissue:

• SNV/indel detection:

  • 18 out of 19 tumor variants were detected in bile cfDNA, achieving a sensitivity of 94.7% and specificity of 99.9%

• CNV detection:

  • 15 out of 20 tumor CNVs were identified in bile cfDNA, with a sensitivity of 75.0% and specificity of 98.9%

• Therapeutic target CNVs identified in bile cfDNA:

  • ERBB2, CDK6, MET, PIK3CA, ROS1, CCNE1, and FLT1

• 13 patients

• AJCC stage I-IV BTCs (7 gallbladder 3 other)

Bile vs. plasma vs. tissue:

• cfDNA yield (n = 11): median 1,918 (bile) vs. 63.1 ng/mL (plasma) (P = 0.0017)

• Detection rate: 84.6% vs. 53.8% vs. 100%

• Median MAF: 1.51–2.68% vs. 1.20% vs. 16.32%

• Top-10 gene concordance: 90% vs. 85% vs. 35%

• 68 patients

• Suspicious biliary strictures

• KRAS: 18 mutations were detected in tissue samples, with 5 additional mutations identified exclusively in bile

• TP53: 13 mutations were detected in tissue samples; 4 additional mutations were found only in bile, while 1 mutation present in tissue was not detected in bile

Superior performance demonstrated in 30 paired bile and tissue samples:

• Sensitivity: 96.4%

• Specificity: 69.2%