The value of metagenomic next-generation sequencing with different nucleic acid extracting methods of cell-free DNA or whole-cell DNA in the diagnosis of non-neutropenic pulmonary aspergillosis

作者：Xiaomin Cai, Chao Sun, Huanhuan Zhong, Y. Cai, Min Cao, Li Wang, Wenkui Sun, Yujian Tao, Guoer Ma, Baoju Huang, Shengmei Yan, Jinjin Zhong, Jiamei Wang, Yajie Lu, Yuanlin Guan, Mengyue Song, Yujie Wang, Yuanyuan Li, Xin Su · 发表于：Frontiers in Cellular and Infection Microbiology · 年份：2024 · DOI：10.3389/fcimb.2024.1398190 · 被引用次数：5 · 研究领域：Antifungal resistance and susceptibility、Actinomycetales infections and treatment、Infectious Diseases and Mycology

Purpose 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. Methods 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. Results 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 Aspergillus detected by BALF-cfDNA, used to predict “True positive” pulmonary aspergillosis patients, was 0.779, with a cut-off value greater than 4.5. Conclusion We propos...