Chamba Clara, Jennings Daisy, Shungu Rehema, Christopher Heavenlight, Josephat Emmanuel, Howard Kieran, Dreau Helene, Burns Adam, Mawalla William, Mapendo Priscus, Mnango Leah, Legason Ismail, Elias Edrick, Achola Caroline, Cutts Anthony, Balandya Emmanuel, Schuh Anna
Muhimbili University of Health and Allied Sciences, Dar es Salaam, Tanzania.
University of Oxford, Oxford, United Kingdom.
JCO Glob Oncol. 2025 Jan;11:e2400210. doi: 10.1200/GO.24.00210. Epub 2025 Jan 9.
Epstein-Barr virus (EBV)-positive Burkitt lymphoma (BL) affects children in sub-Saharan Africa, but diagnosis via tissue biopsy is challenging. We explored a liquid biopsy approach using targeted next-generation sequencing to detect the -immunoglobulin (-Ig) translocation and EBV DNA, assessing its potential for minimally invasive BL diagnosis.
The panel included targets for the characteristic -Ig translocation, mutations in intron 1 of , mutations in exon 2 of , and three EBV genes: EBV-encoded RNA (EBER)1, EBER2, and EBV nuclear antigen 2. It was first tested in a small derivation cohort of four precharacterized BL-derived cell lines with known translocation status and eight precharacterized plasma samples with known EBV DNA status by quantitative polymerase chain reaction (qPCR). These different data modalities were combined to assess the accuracy of this approach in the diagnosis of BL in 20 patient plasma samples in Tanzania and Uganda.
The next-generation sequencing panel detected three of four -Ig translocations in the BL-derived cell lines. EBV viral load by targeted sequencing correlated strongly with qPCR results (Spearman's rho = 0.94) in precharacterized plasma samples. Using the patient plasma samples, mutations in intron 1 were associated with the presence of a translocation with 25 or more mutations being predictive of a translocation with AUC, sensitivity, and specificity of 1. Overall, liquid biopsy parameters associated with a diagnosis of BL ( < .05) included cell-free DNA concentration, circulating tumor DNA concentration, intron 1 mutations, -Ig translocation, and autosome entropy. Integrating these parameters into a diagnostic model demonstrated excellent performance with an AUC of 0.95, sensitivity of 0.9, and specificity of 1.
This analysis demonstrates the potential of liquid biopsy to improve BL diagnosis in settings with limited pathology resources. Validation of our approach in a larger data set is needed.
爱泼斯坦-巴尔病毒（EBV）阳性的伯基特淋巴瘤（BL）影响撒哈拉以南非洲地区的儿童，但通过组织活检进行诊断具有挑战性。我们探索了一种液体活检方法，使用靶向新一代测序来检测免疫球蛋白（-Ig）易位和EBV DNA，评估其在微创性BL诊断中的潜力。
该检测 panel 包括特征性-Ig易位的靶点、基因内含子1的突变、基因外显子2的突变以及三个EBV基因：EBV编码RNA（EBER）1、EBER2和EBV核抗原2。首先在一个小型衍生队列中进行测试，该队列包括四个预先特征化的具有已知易位状态的BL衍生细胞系和八个预先特征化的通过定量聚合酶链反应（qPCR）具有已知EBV DNA状态的血浆样本。将这些不同的数据模式结合起来，以评估该方法在坦桑尼亚和乌干达的20例患者血浆样本中诊断BL的准确性。
新一代测序检测 panel 在BL衍生细胞系中检测到四个-Ig易位中的三个。在预先特征化的血浆样本中，通过靶向测序得到的EBV病毒载量与qPCR结果高度相关（斯皮尔曼相关系数=0.94）。使用患者血浆样本，基因内含子1的突变与易位的存在相关，25个或更多突变可预测易位，其曲线下面积（AUC）、敏感性和特异性均为1。总体而言，与BL诊断相关的液体活检参数（P<0.）包括游离DNA浓度、循环肿瘤DNA浓度、基因内含子1突变、-Ig易位和常染色体熵。将这些参数整合到一个诊断模型中显示出优异的性能，AUC为0.95，敏感性为0.9，特异性为1。
该分析证明了液体活检在病理资源有限的情况下改善BL诊断的潜力。需要在更大的数据集中对我们的方法进行验证。
