Department of Respiratory and Critical Care Medicine, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, China.
Department of Respiratory and Critical Medicine, Jinling Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, China.
Front Cell Infect Microbiol. 2024 Jul 29;14:1398190. doi: 10.3389/fcimb.2024.1398190. eCollection 2024.
Metagenomic next-generation sequencing(mNGS) is a novel molecular diagnostic technique. For nucleic acid extraction methods, both whole-cell DNA (wcDNA) and cell-free DNA (cfDNA) are widely applied with the sample of bronchoalveolar lavage fluid (BALF). We aim to evaluate the clinical value of mNGS with cfDNA and mNGS with wcDNA for the detection of BALF pathogens in non-neutropenic pulmonary aspergillosis.
mNGS with BALF-cfDNA, BALF-wcDNA and conventional microbiological tests (CMTs) were performed in suspected non-neutropenic pulmonary aspergillosis. The diagnostic value of different assays for pulmonary aspergillosis was compared.
BALF-mNGS (cfDNA, wcDNA) outperformed CMTs in terms of microorganisms detection. Receiver operating characteristic (ROC) analysis indicated BALF-mNGS (cfDNA, wcDNA) was superior to culture and BALF-GM. Combination diagnosis of either positive for BALF-mNGS (cfDNA, wcDNA) or CMTs is more sensitive than CMTs alone in the diagnosis of pulmonary aspergillosis (BALF-cfDNA+CMTs/BALF-wcDNA+CMTs vs. CMTs: ROC analysis: 0.813 vs.0.66, P=0.0142/0.796 vs.0.66, P=0.0244; Sensitivity: 89.47% vs. 47.37%, P=0.008/84.21% vs. 47.37%, P=0.016). BALF-cfDNA showed a significantly greater reads per million (RPM) than BALF-wcDNA. The area under the ROC curve (AUC) for RPM of detected by BALF-cfDNA, used to predict "True positive" pulmonary aspergillosis patients, was 0.779, with a cut-off value greater than 4.5.
We propose that the incorporation of BALF-mNGS (cfDNA, wcDNA) with CMTs improves diagnostic precision in the identification of non-neutropenic pulmonary aspergillosis when compared to CMTs alone. BALF-cfDNA outperforms BALF-wcDNA in clinical value.
宏基因组下一代测序（mNGS）是一种新型的分子诊断技术。对于核酸提取方法，支气管肺泡灌洗液（BALF）的样本既可以应用全细胞 DNA（wcDNA）也可以应用游离细胞 DNA（cfDNA）。我们旨在评估 cfDNA 和 wcDNA 的 mNGS 检测非中性粒细胞减少性肺曲霉病中 BALF 病原体的临床价值。
对疑似非中性粒细胞减少性肺曲霉病进行 mNGS 检测 BALF-cfDNA、BALF-wcDNA 和常规微生物学检测（CMTs）。比较不同检测方法对肺曲霉病的诊断价值。
BALF-mNGS（cfDNA、wcDNA）在微生物检测方面优于 CMTs。受试者工作特征（ROC）分析表明，BALF-mNGS（cfDNA、wcDNA）优于培养和 BALF-GM。BALF-mNGS（cfDNA、wcDNA）或 CMTs 联合诊断比 CMTs 单独诊断肺曲霉病更敏感（BALF-cfDNA+CMTs/BALF-wcDNA+CMTs 与 CMTs 的 ROC 分析：0.813 与 0.66，P=0.0142/0.796 与 0.66，P=0.0244；敏感性：89.47%与 47.37%，P=0.008/84.21%与 47.37%，P=0.016）。BALF-cfDNA 的每百万读数（RPM）明显高于 BALF-wcDNA。BALF-cfDNA 检测到的 RPM 的 ROC 曲线下面积（AUC），用于预测“真阳性”肺曲霉病患者，为 0.779，截断值大于 4.5。
我们提出，与 CMTs 相比，将 BALF-mNGS（cfDNA、wcDNA）与 CMTs 联合应用可提高非中性粒细胞减少性肺曲霉病的诊断精度。BALF-cfDNA 在临床价值方面优于 BALF-wcDNA。
