Circulating plasma EBV DNA: a potential tool to facilitate diagnosis of pediatric Burkitt lymphoma in sub-Saharan Africa

Abstract: In sub-Saharan Africa (SSA), many pediatric patients with Burkitt lymphoma (BL) perish because of diagnostic delays. About 95% of pediatric BL cases in SSA are Epstein-Barr virus (EBV) positive. We assessed plasma EBV DNA in 400 pediatric patients with BL and 400 controls, frequency-matched on sex, age, and country, enrolled in the Epidemiology of Burkitt Lymphoma in East African Children and Minors study in Uganda, Kenya, and Tanzania (2010-2016). EBV was measured using a digital droplet polymerase chain reaction assay targeting EBV BamHI-W internal repeats, duplexed with RPP30 human housekeeping gene. The study population was predominantly male (63% of cases of BL, and 64% of controls), with an average age of ∼7.5 years. EBV DNA was detected in 309 (77.3%) cases of BL, and 62 (15.5%) controls. The mean plasma EBV DNA levels were 5.00 (standard deviation [SD] 1.63) log10 copies per mL for cases with BL vs 1.94 (SD 1.35) for controls (P < .0001). Excluding 50 cases with BL and 61 controls with indeterminate (RPP30 and EBV negative) samples, the maximum sensitivity of plasma EBV DNA detection to discriminate cases with BL from controls was 88.3% (95% confidence interval [CI], 84.4-91.5), with 81.7% (95% CI, 77.2-85.7) specificity, and area under the curve 91.9% (95% CI, 89.7-93.9). A specificity of 100% was achieved at a threshold of 4.19 log10 EBV copies per mL, which reduced sensitivity to 66.6%. Assay accuracy varied from 83% to 87.4% at different thresholds. Testing for EBV DNA could facilitate the recognition of BL earlier in SSA, a critical step in improving BL cure rates in the region.