Blind Brush Biopsy: Quantification of Epstein-Barr Virus and Its Host DNA Methylation in the Detection of Nasopharyngeal Carcinoma

Caoli Tang; Xizhao Li; Yumeng Zhang; Ting Zhou; Xiaojing Yang; Ying Liao; Tongmin Wang; Yongqiao He; Wenqiong Xue; Weihua Jia; Xiaohui Zheng

doi:10.34133/research.0475

Blind Brush Biopsy: Quantification of Epstein-Barr Virus and Its Host DNA Methylation in the Detection of Nasopharyngeal Carcinoma

Research (Wash D C). 2024 Sep 24:7:0475. doi: 10.34133/research.0475. eCollection 2024.

Authors

Caoli Tang^{1

2}, Xizhao Li¹, Yumeng Zhang^{1

2}, Ting Zhou¹, Xiaojing Yang¹, Ying Liao¹, Tongmin Wang¹, Yongqiao He¹, Wenqiong Xue¹, Weihua Jia^{1

2}, Xiaohui Zheng¹

Affiliations

¹ State Key Laboratory of Oncology in South China, Guangdong Key Laboratory of Nasopharyngeal Carcinoma Diagnosis and Therapy, Guangdong Provincial Clinical Research Center for Cancer, Sun Yat-sen University Cancer Center, Guangzhou 510060, P. R. China.
² School of Public Health, Sun Yat-sen University, Guangzhou 510080, P. R. China.

Abstract

Background: The nasopharyngeal brush sampling can effectively collect samples from the nasopharynx. The blind brush sampling does not require the guidance of endoscopy, which is favorable for implementation and dissemination in the community. This study explored methylation markers for nasopharyngeal carcinoma (NPC) at both Epstein-Barr virus (EBV) and its host genome levels, aiming to construct a blind brushing diagnostic method. Methods: EBV DNA capture and methylation sequencing and GEO Illumina 450K methylation array data were used respectively for the discovery of EBV and host methylation markers. The diagnostic method was built in training cohort (n = 347) and validated in an independent validation cohort (n = 155). Results: A total of 1 EBV methylation marker (BILF2) and 6 host methylation markers (ITGA4, IMPA2, ITPKB, PI9, AMIGO2, and VAV3) were identified. Both EBV and host methylation markers were almost exclusively detected in NPC samples, with negligible detection in control samples. In validation cohort, the diagnostic method that included only the EBV BILF2 marker showed a sensitivity and specificity of 80.22% and 98.44%, respectively. When combining the EBV-derived marker BILF2 with the host-derived marker IMPA2, the diagnostic method's sensitivity increased to 84.62%, while the specificity remained unchanged (IDI = 4.4%, P = 0.0419). Conclusion: Overall, the blind nasopharyngeal brushing diagnostic method, combining EBV and host methylation markers, showed great potential in NPC detection and could promote its application in nonclinical screening of NPC.