目的：本研究拟探讨基于超多重聚合酶链反应（polymerase chain reaction，PCR）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。 方法：本研究共纳入2023年5-7月期间209例临床怀疑IPA患者，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、革兰染色涂片检测。此外，我们还分析了2023年本检验中心共计13452份BALF样本的tNGS检出数据。 结果：在209例疑似IPA患者送检的BALF样本，tNGS在183例中检出曲霉菌，阳性率达到87.6%。烟曲霉是检出率最高的种属（127例，69.4%），其次为黄曲霉（43例，23.5%）、土曲霉（7例，3.83%）和黑曲霉（6例，3.28%）。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%，低于tNGS技术（P＜0.001），但均显著高于革兰染色涂片（20.1%）。在183例tNGS曲霉菌检出阳性的样本中，168例存在其它病原体检出，占比91.8%。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，在本检验中心13452份BALF样本中，tNGS在2131份（13.7%）样本中检出曲霉菌。烟曲霉（56.9%）是最常检出的种属，其次为黄曲霉（33.4%）、黑曲霉（3.5%）和土曲霉（1.7%），两种以上曲霉菌种属检出占比为4.5%。曲霉菌合并病原体检出占比91.1%，其中，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%）、结核分枝杆菌复合群（6.76%）是曲霉菌合并检出较高的病原体。 结论：tNGS技术在IPA诊断中较GM试验有着更高的敏感性和特异性。此外，tNGS技术还能够准确分型曲霉菌种属，且对于真菌混合感染的诊断具有优势。

Objective: This study investigates the role of multiplex PCR -based targeted next-generation sequencing (tNGS) in the diagnosis of invasive pulmonary aspergillosis (IPA). Methods: A total of 209 patients with clinical suspicion of IPA were included in this study from January to July 2023. Each patient underwent fiber bronchoscopy, and bronchoalveolar lavage fluid (BALF) was collected for analysis. tNGS, galactomannan (GM) testing, and Gram staining were performed on these samples. Additionally, we analyzed tNGS detection data from 13,452 BALF samples collected at our test center throughout 2023.. Results: Among the 209 BALF samples from suspected IPA patients, tNGS detected Aspergillus in 183 cases, yielding a positive rate of 87.6%. The species distribution revealed that aspergillus fumigatus had the highest detection rate (127 cases, 69.4%), followed by Aspergillus aflatus (43 cases, 23.5%), Aspergillus terreus (7 cases, 3.83%) and Aspergillus Niger (6 cases, 3.28%). Using a GM test threshold of ≥1.0, the detection rate of aspergillus was 70.3%, significantly lower than that of tNGS (P < 0.001), but notably higher than that from Gram staining smear (20.1%). Among the 183 tNGS positive samples, additional pathogens were detected in 168 cases, accounting for 91.8%. The sequence number of tNGS in patients with positive GM test was significantly higher than that in patients with negative GM test (P < 0.001). In addition, among the 13452 BALF samples, tNGS detected aspergillus in 2131 (13.7%) samples. Aspergillus fumigatus (56.9%) was the most commonly detected species, followed by Aspergillus aflatus (33.4%), Aspergillus Niger (3.5%) and Aspergillus terreus (1.7%), and more than two aspergillus species accounted for 4.5%. A total of 91.1% of the pathogens detected were associated with Aspergillus. awith the most common being (25.9%), Klebsiella pneumoniae (14.2%), Acinetobacter baumannii (12.6%), novel coronavirus (12.1%), Pseudomonas aeruginosa (9.53%) and Mycobacterium tuberculosis complex group (6.76%) . Conclusions:The tNGS technique has higher sensitivity and specificity than GM test in IPA diagnosis. Moreover, tNGS enables accurate identification of Aspergillus species and offers significant advantages in diagnosing mixed fungal infections..