本研究旨在探讨基于超多重聚合酶链反应(polymerase chain reaction，PCR)的靶向下一代测序(targeted next-generation sequencing, tNGS)技术在侵袭性肺曲霉菌病(invasive pulmonary aspergillosis，IPA)诊断中的价值。本研究共纳入2023年5月—7月期间209例临床怀疑IPA患者，通过纤维支气管镜检查取肺泡灌洗液(bronchoalveolar lavage fluid，BALF)，分别行tNGS、半乳甘露聚糖(glactomannan, GM)试验、革兰氏染色涂片检测。在疑似IPA患者的BALF样本中，tNGS检出183例曲霉菌，阳性率达87.6%。烟曲霉是检出率最高的种属(69.4%)，其次为黄曲霉(23.5%)、土曲霉(3.8%)和黑曲霉(3.3%)。以BALF样本GM试验≥1.0为标准，GM试验对于曲霉菌的检出率为70.3%，检出率低于tNGS技术(P<0.001)，但均显著高于革兰氏染色涂片(20.1%)。在183例tNGS检出曲霉菌的样本中，168例(91.8%)检出其他病原体。GM试验阳性的患者，tNGS曲霉菌的序列数显著高于GM试验阴性的患者(P<0.001)。此外，本检验中心2023年收集的15 583 份BALF样本中，tNGS在2 131 份(13.7%)样本中检出曲霉菌。其中，曲霉菌合并其他病原体检出占比91.1%，流感病毒(25.9%)、肺炎克雷伯菌(14.2%)、鲍曼不动杆菌(12.6%)、新型冠状病毒(12.1%)、铜绿假单胞菌(9.53%)是合并检出率较高的病原体。本研究证实，tNGS技术在IPA诊断中较GM试验的敏感性和特异性更高，且能准确对曲霉菌进行分型，对真菌混合感染诊断有优势。

This study evaluated the diagnostic utility of ultra-multiplex polymerase chain reaction(PCR)-based targeted next-generation sequencing (tNGS) in invasive pulmonary aspergillosis (IPA). A cohort of 209 patients clinically suspected of IPA during May and July 2023 underwent bronchoalveolar lavage fluid (BALF) collection via fiberoptic bronchoscopy. Samples were analyzed using tNGS, galactomannan (GM) assay, and Gram-stained smear. Among 209 suspected IPA cases, tNGS identified Aspergillus in 183 samples, achieving an 87.6% positivity rate. Aspergillus fumigatus was the predominant species (69.4%), followed by Aspergillus flavus (23.5%), Aspergillus terreus (3.83%), and Aspergillus niger (3.3%). GM assay, using a BALF GM index cutoff of ≥1.0, yielded a significantly lower positivity rate of 70.3% (P<0.001), but both outperformed Gram-stained smear (20.1%). Co-detection of other pathogens occurred in 168 tNGS-positive samples (91.8%). Notably, tNGS-derived Aspergillus read counts were significantly higher in GM-positive versus GM-negative patients (P<0.001). Among 15 583 BALF samples analyzed in 2023, tNGS identified Aspergillus in 2 131 (13.7%) samples. Co-pathogens were detected in 91.1% of Aspergillus-positive cases, with influenza virus (25.9%), Klebsiella pneumoniae (14.2%), Acinetobacter baumannii (12.6%), SARS-CoV-2 (12.1%), and Pseudomonas aeruginosa (9.53%) being the predominant other pathogens. This study demonstrates that tNGS surpasses the GM assay in both sensitivity and specificity for IPA diagnosis, enables precise Aspergillus species identification, and excels in detecting fungal co-infections.