Wang Hansheng, Chen Xiao, You Hui, Wang Yunyun, Xia Xianru, Tang Yijun, Ren Tao, Wang Meifang
Department of Pulmonary and Critical Care Medicine, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei, China.
Department of Laboratory, Shiyan Maternal and Child Health Hospital, Hubei University of Medicine, Shiyan, Hubei, China.
J Infect Dis. 2025 Jul 11;231(6):1609-1618. doi: 10.1093/infdis/jiaf044.
Multiplex polymerase chain reaction (PCR)-based targeted next-generation sequencing (tNGS) is a promising tool for distinguishing lower respiratory tract infections in clinical practice, and its detectable pathogen spectrum can cover more than 95% of clinical cases, but there is limited information on systematic evaluation of the clinical use of multiplex PCR-based tNGS (mp-tNGS) in invasive pulmonary aspergillosis (IPA) cases. We aim to assess mp-tNGS in bronchoalveolar lavage fluid (BALF) for Aspergillus detection in patients with suspected IPA to provide a reliable basis for initiating antifungal therapy without microbiological or histopathological evidence.
We prospectively enrolled a cohort of consecutive patients with suspected IPA; all had undergone serum/BALF galactomannan antigen (GM), BALF mp-tNGS, and traditional tests (direct smear and culture of respiratory specimens). EORTC/MSG and FUDICU criteria or clinical compound diagnosis were used for IPA diagnosis.
Thirty-two patients were diagnosed with IPA and 42 with non-IPA. Compared with the final diagnosis, the sensitivity of BALF mp-tNGS was 87.5%, while the sensitivities of traditional tests, serum GM, and BALF GM assay were 43.8%, 21.9%, and 62.5%, respectively. The specificity of BALF mp-tNGS was 90.5%, which was similar to traditional tests. The average turnaround time for Aspergillus detection by BALF mp-tNGS was 22.10 hours (SD 2.49 hours), which was significantly faster than traditional tests.
BALF mp-tNGS showed good performance in identification of Aspergillus in nonneutropenic IPA patients. Importantly, positive mp-tNGS in BALF can provide a basis for early antifungal therapy before microbiological evidence is available.
基于多重聚合酶链反应（PCR）的靶向二代测序（tNGS）是临床实践中区分下呼吸道感染的一种有前景的工具，其可检测的病原体谱能覆盖超过95%的临床病例，但关于基于多重PCR的tNGS（mp-tNGS）在侵袭性肺曲霉病（IPA）病例中临床应用的系统评价信息有限。我们旨在评估支气管肺泡灌洗液（BALF）中的mp-tNGS用于疑似IPA患者曲霉菌检测的情况，以便在缺乏微生物学或组织病理学证据时为启动抗真菌治疗提供可靠依据。
我们前瞻性纳入了一组连续的疑似IPA患者队列；所有患者均接受了血清/BALF半乳甘露聚糖抗原（GM）检测、BALF mp-tNGS检测以及传统检测（呼吸道标本直接涂片和培养）。采用欧洲癌症研究与治疗组织/侵袭性真菌感染协作组和美国国立过敏和传染病研究所（EORTC/MSG）标准或临床复合诊断法对IPA进行诊断。
32例患者被诊断为IPA，42例为非IPA。与最终诊断相比，BALF mp-tNGS的敏感性为87.5%，而传统检测、血清GM检测和BALF GM检测的敏感性分别为43.8%、21.9%和62.5%。BALF mp-tNGS的特异性为90.5%，与传统检测相似。BALF mp-tNGS检测曲霉菌的平均周转时间为22.10小时（标准差2.49小时），明显快于传统检测。
BALF mp-tNGS在非中性粒细胞减少的IPA患者曲霉菌鉴定中表现良好。重要的是，BALF中mp-tNGS呈阳性可为在获得微生物学证据之前尽早开始抗真菌治疗提供依据。
