血浆宏基因组下一代测序与聚合酶链反应方法在鼻咽癌患者中监测爱泼斯坦-巴尔病毒载量的比较

Comparison of Plasma Metagenomic Next-generation Sequencing and PCR Methods for Epstein-Barr Virus Viral Load Monitoring in Nasopharyngeal Carcinoma.

作者信息

Vasudevan Harish N, Lazar Ann A, Reyes Kevin, Rong D A, Arevalo Shaun, Federman Scot, Chan Jason W, Chiu Charles Y, Miller Steve, Yom Sue S

机构信息

Department of Radiation Oncology, University of California, San Francisco, CA, U.S.A.

Division of Oral Epidemiology and Division of Biostatistics, University of California, San Francisco, CA, U.S.A.

出版信息

Anticancer Res. 2024 Dec;44(12):5445-5453. doi: 10.21873/anticanres.17370.

DOI:10.21873/anticanres.17370

PMID:39626922

Abstract

BACKGROUND/AIM: Plasma Epstein-Barr virus (EBV) viral load measurement is prognostic in nasopharyngeal carcinoma (NPC) disease monitoring; however, a consensus measurement approach does not exist. This study characterized the clinical performance of metagenomic next-generation sequencing (mNGS), an unbiased sequencing-based assay distinct from polymerase chain reaction (PCR) or targeted sequencing approaches, in 73 peripheral blood specimens from 32 patients diagnosed with NPC.

PATIENTS AND METHODS

Samples were analyzed for plasma EBV viral load either by mNGS profiling or PCR-based assays (either LMP2 or BAMHI-W PCR) and compared to tumor presence by clinical assessment. Plasma mNGS-based EBV detection was quantified as reads per million (RPM).

RESULTS

Plasma mNGS displayed similar overall performance (100% sensitivity, 86% specificity, 92% accuracy) to BAMHI-W PCR (100% sensitivity, 86% specificity, 94% accuracy) and superior performance to the LMP2 PCR assay (36% sensitivity, 56% specificity, 45% accuracy). In a subset of 13 patients who underwent longitudinal analysis, plasma mNGS EBV RPM correlated with cancer recurrence (95%CI Pre-CRT=232.10±214; 95%CI Post-CRT=0.34±0.32; 95%CI difference=-231.70±214; *p=0.03, paired t-test), suggesting plasma mNGS exhibits potential for monitoring recurrence.

CONCLUSION

Plasma mNGS is a distinct method for EBV titer measurement in NPC patients and more broadly, is a promising method for non-invasive monitoring of disease status for infection-associated malignancies.

摘要

背景/目的：血浆爱泼斯坦-巴尔病毒（EBV）载量检测在鼻咽癌（NPC）疾病监测中具有预后价值；然而，目前尚无统一的检测方法。本研究对32例确诊为NPC的患者的73份外周血标本进行宏基因组下一代测序（mNGS），这是一种基于测序的无偏检测方法，与聚合酶链反应（PCR）或靶向测序方法不同，旨在分析其临床性能。

患者与方法

通过mNGS分析或基于PCR的检测方法（LMP2或BAMHI-W PCR）对样本进行血浆EBV载量分析，并与临床评估的肿瘤存在情况进行比较。基于血浆mNGS的EBV检测定量为每百万读数（RPM）。

结果

血浆mNGS的总体性能（敏感性100%，特异性86%，准确性92%）与BAMHI-W PCR（敏感性100%，特异性86%，准确性94%）相似，且性能优于LMP2 PCR检测（敏感性36%，特异性56%，准确性45%）。在13例接受纵向分析的患者亚组中，血浆mNGS EBV RPM与癌症复发相关（95%CI 放疗前=232.10±214；95%CI 放疗后=0.34±0.32；95%CI 差异=-231.70±214；*p=0.03，配对t检验），提示血浆mNGS在监测复发方面具有潜力。