Sequencing-based counting and size profiling of plasma Epstein–Barr virus DNA enhance population screening of nasopharyngeal carcinoma

Significance We identified differentiating molecular characteristics of plasma EBV DNA between nasopharyngeal carcinoma (NPC) patients and non-NPC subjects. Sequencing-based analysis revealed higher amounts of plasma EBV DNA and generally longer fragment lengths of plasma viral molecules in NPC patients than in non-NPC subjects. Based on these findings, we have developed a highly accurate blood-based test for screening of NPC. Such an approach is shown to enhance the positive predictive value and demonstrate a superior performance for NPC screening. It also obviates the need of a follow-up blood sample and therefore allows single time-point testing. We believe that this more clinically practical protocol would facilitate NPC screening on a population scale.