Department of Clinical Laboratory, Liaocheng People's Hospital, Liaocheng, China.
Department of Central Laboratory, Liaocheng People's Hospital, Liaocheng, China.
Clin Chim Acta. 2024 Apr 15;557:117874. doi: 10.1016/j.cca.2024.117874. Epub 2024 Mar 12.
Metagenomic next-generation sequencing (mNGS) is valuable for pathogen identification; however, distinguishing between infectious diseases and conditions with potentially similar clinical manifestations, including malignant tumors, is challenging. Therefore, we developed a method for simultaneous detection of infectious pathogens and cancer in blood samples.
Plasma samples (n = 244) were collected from 150 and 94 patients with infections and hematological malignancies, respectively, and analyzed by mNGS for pathogen detection, alongside human tumor chromosomal copy number variation (CNV) analysis (≥5Mbp or 10Mbp CNV region). Further, an evaluation set, comprising 87 plasma samples, was analyzed by mNGS and human CNV analysis, to validate the feasibility of the method.
Among 94 patients with hematological malignancy, sensitivity values of CNV detection for tumor diagnosis were 69.15 % and 32.98 % for CNV region 5Mbp and 10Mbp, respectively, with corresponding specificities of 92.62 % and 100 % in the infection group. Area under the ROC curve (AUC) values for 5Mbp and 10Mbp region were 0.825 and 0.665, respectively, which was a significant difference of 0.160 (95 % CI: 0.110-0.210; p < 0.001), highlighting the superiority of 5Mbp output region data. Six patients with high-risk CNV results were identified in the validation study: three with history of tumor treatment, two eventually newly-diagnosed with hematological malignancies, and one with indeterminate final diagnosis.
Concurrent CNV analysis alongside mNGS for infection diagnosis is promising for detecting malignant tumors. We recommend adopting a CNV region of 10Mbp over 5Mbp for our model, because of the lower false-positive rate (FPR).
宏基因组下一代测序（mNGS）在病原体鉴定方面具有重要价值；然而，区分具有相似临床表现的传染病和病症（包括恶性肿瘤）具有挑战性。因此，我们开发了一种同时检测血液样本中感染性病原体和癌症的方法。
从 150 例感染患者和 94 例血液恶性肿瘤患者中分别采集血浆样本（n=244），通过 mNGS 进行病原体检测，同时进行人类肿瘤染色体拷贝数变异（CNV）分析（≥5Mbp 或 10Mbp CNV 区域）。此外，还对包含 87 个血浆样本的验证集进行了 mNGS 和人类 CNV 分析，以验证该方法的可行性。
在 94 例血液恶性肿瘤患者中，CNV 检测对肿瘤诊断的灵敏度分别为 69.15%和 32.98%，对于 5Mbp 和 10Mbp 区域的 CNV 区域，在感染组中，相应的特异性分别为 92.62%和 100%。5Mbp 和 10Mbp 区域的 ROC 曲线下面积（AUC）值分别为 0.825 和 0.665，差异具有统计学意义（95%CI：0.110-0.210；p<0.001），这表明 5Mbp 输出区域数据具有优势。在验证研究中发现了 6 例具有高危 CNV 结果的患者：3 例有肿瘤治疗史，2 例最终新诊断为血液恶性肿瘤，1 例最终诊断不确定。
在感染诊断中，同时进行 CNV 分析和 mNGS 检测有望用于检测恶性肿瘤。我们建议采用 10Mbp 的 CNV 区域而不是 5Mbp，因为它的假阳性率（FPR）较低。
