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ORIGINAL RESEARCH article

Front. Cell. Infect. Microbiol.
Sec. Clinical Infectious Diseases
Volume 15 - 2025 | doi: 10.3389/fcimb.2025.1474083
This article is part of the Research Topic Targeted Next-Generation Sequencing for Pathogen and Antimicrobial Resistance (AMR) Identification and Profiling View all 8 articles

Performance of multiplex PCR-based targeted next-generation sequencing in bronchoalveolar lavage fluid for the diagnosis of invasive pulmonary aspergillosis in non-neutropenic patients

Provisionally accepted
Hansheng Wang Hansheng Wang 1Xiao Chen Xiao Chen 2*Wenya Han Wenya Han 1Yuquan Liu Yuquan Liu 1*Hui You Hui You 1*Xia Xianru Xia Xianru 1Dan Yu Dan Yu 1*Yijun Tang Yijun Tang 1*Meifang Wang Meifang Wang 1*Tao Ren Tao Ren 1*
  • 1 Taihe Hospital, Hubei University of Medicine, Shiyan, China
  • 2 Shiyan Maternity and Child Health Care Hospital, Shiyan, China

The final, formatted version of the article will be published soon.

    The diagnosis of invasive pulmonary aspergillosis (IPA) depends on histopathology and mycological culture; rapid and reliable diagnosis of IPA is crucial. Multiplex polymerase chain reaction (PCR)-based Targeted targeted next-generation sequencing (tNGS) is a 2 promising tool for distinguishing lower respiratory tract infections (LRTIs) in clinical practiceidentifying pathogens, 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 IPA cases. We aim to assess mp-tNGS in bronchoalveolar lavage fluid (BALF) for Aspergillus detection in suspected IPA patients, and to provide a reliable basis for initiating antifungal therapy without microbiological or histopathological evidence.We prospectively enrolled a cohort of consecutive patients suspected of IPA, all of whomthem had received undergone serum/BALF galactomannan antigen (GM), BALF mp-tNGS, and traditional tests (direct smear and culture of respiratory specimens), and EORTC/MSGERC criteria were used for IPA diagnosis. Results: Seventy-four patients with suspected IPA were eventually included in our study. 32 Thirty-two patients were diagnosed with IPA and 42 with non-IPA. The sensitivity and specificity of BALF mp-tNGS for Aspergillus were calculated, including the diagnostic turnaround time (TAT). Compared with the final diagnosis, the sensitivity of BALF mp-tNGS was 87.5%, while the sensitivity of traditional tests, serum GM and BALF GM assay was 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 TAT for Aspergillus detection by BALF mp-tNGS was 22.10±2.49h, which was significantly faster than that by traditional tests. Conclusion: BALF mp-tNGS showed good performance in identification of Aspergillus in non-neutropenic IPA patients.Importantly, positive mp-tNGS in BALF can provide a basis for early antifungal therapy before microbiological evidence is available.

    Keywords: Multiplex PCR-based targeted next generation sequencing (mp-tNGS), Bronchoalveolar lavage fluid (BALF), Invasive pulmonary aspergillosis (IPA), histopathology, galactomannan (GM), mycological culture

    Received: 01 Aug 2024; Accepted: 02 Jan 2025.

    Copyright: © 2025 Wang, Chen, Han, Liu, You, Xianru, Yu, Tang, Wang and Ren. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence:
    Xiao Chen, Shiyan Maternity and Child Health Care Hospital, Shiyan, 442000, China
    Yuquan Liu, Taihe Hospital, Hubei University of Medicine, Shiyan, China
    Hui You, Taihe Hospital, Hubei University of Medicine, Shiyan, China
    Dan Yu, Taihe Hospital, Hubei University of Medicine, Shiyan, China
    Yijun Tang, Taihe Hospital, Hubei University of Medicine, Shiyan, China
    Meifang Wang, Taihe Hospital, Hubei University of Medicine, Shiyan, China
    Tao Ren, Taihe Hospital, Hubei University of Medicine, Shiyan, China

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