Combined multiplex polymerase chain reaction-based targeted next-generation sequencing and serum 1, 3-β-D-glucan for differential diagnosis of Pneumocystis pneumonia and Pneumocystis colonization.

IF 4.8 2区医学 Q2 IMMUNOLOGY

Frontiers in Cellular and Infection Microbiology Pub Date : 2025-09-18 eCollection Date: 2025-01-01 DOI:10.3389/fcimb.2025.1611391

Hansheng Wang, Xiao Chen, Xiaofeng Wu, Qizhen Cao, Yi Wu, Fang Wang, Yunyun Wang, Yanhui Zhou, Yijun Tang, Tao Ren, Meifang Wang

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引用次数: 0

Abstract

Background and objective: Pneumocystis jirovecii pneumonia (PjP) remains an important cause of morbimortality worldwide, and differentiating Pneumocystis jirovecii (P. jirovecii) infection from P. jirovecii colonization (PjC) is crucial for guiding treatment strategies. Multiplex polymerase chain reaction-based targeted next-generation sequencing (mp-tNGS) is a promising tool for identifying lower respiratory tract infections, with a detectable pathogen spectrum that covers more than 95% of clinical infectious cases. This study evaluated mp-tNGS for P. jirovecii identification in bronchoalveolar lavage fluid (BALF) samples combined with serum 1,3-β-D-glucan (BDG) level detection to differentiate PjP and PjC.

Methods: A total of 73 patients were enrolled and the final diagnosis was used as a reference criterion, and patients were divided into the PjP group and PjC group. The clinical data and detection performance of mp-tNGS/serum BDG were analyzed.

Results: The median fungal reads (normalized sequence counts) detected by mp-tNGS were 1522.00 (interquartile range [IQR], 581.5, 4898.0) in the PjP group versus 117.00 (IQR, 79.00, 257.00) in the PjC group (p <0.0001). Correspondingly, BDG levels were 122.5 (88.75,239.3) pg/ml in PjP patients compared to 59.00 (51.0,79.0) pg/ml in PjC patients (p <0.0001). Area under the receiver operator characteristic curve (AUROC) for discriminating PjP from colonization was 0.935 (95% CI: 0.88-0.99) for BALF mp-tNGS and 0.822 (95% CI: 0.72-0.93) for serum BDG. The optimal diagnostic thresholds were determined to be 355 reads for mp-tNGS (sensitivity: 89.1%; specificity: 85.2%) and 84.5 pg/ml for BDG (sensitivity: 85.2%; specificity: 80.4%).

Conclusion: BALF mp-tNGS and serum BDG serve as valuable adjunct diagnostic tools, providing reliable differentiation between P. jirovecii colonization and active infection.

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基于多重聚合酶链反应的新一代靶向测序和血清1,3 -β- d -葡聚糖对肺囊虫肺炎和肺囊虫定植的鉴别诊断

背景与目的：耶氏肺囊虫肺炎（PjP）仍然是世界范围内重要的致死率，区分耶氏肺囊虫感染和耶氏肺囊虫定植（PjC）对指导治疗策略至关重要。基于多重聚合酶链反应的靶向下一代测序（mp-tNGS）是一种很有前途的下呼吸道感染鉴定工具，其可检测的病原体谱覆盖了95%以上的临床感染病例。本研究评价mp-tNGS联合血清1,3-β- d -葡聚糖（BDG）水平检测在支气管肺泡灌洗液（BALF）样品中鉴定吉罗氏假单胞菌的作用，以鉴别PjP和PjC。方法：共纳入73例患者，以最终诊断为参考标准，分为PjP组和PjC组。分析mp-tNGS/血清BDG的临床资料及检测性能。结果：mp-tNGS检测到的真菌reads（归一化序列计数）中位数在PjP组为1522.00（四分位数间距[IQR]， 581.5, 4898.0），而PjC组为117.00（四分位数间距[IQR]， 79.00, 257.00）（p 0.0001）。相应的，PjP患者的BDG水平为122.5 (88.75,239.3)pg/ml，而PjC患者的BDG水平为59.00 (51.0,79.0)pg/ml （p 0.0001）。BALF mp-tNGS区分PjP和定植的受试者操作特征曲线下面积（AUROC）为0.935 (95% CI: 0.88-0.99)，血清BDG为0.822 （95% CI: 0.72-0.93）。最佳诊断阈值确定为mp-tNGS为355 reads（敏感性：89.1%，特异性：85.2%），BDG为84.5 pg/ml（敏感性：85.2%，特异性：80.4%）。结论：BALF mp-tNGS和血清BDG可作为有价值的辅助诊断工具，为区分氏弓形虫定殖和活动性感染提供可靠依据。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Frontiers in Cellular and Infection Microbiology IMMUNOLOGY-MICROBIOLOGY

CiteScore

7.90

自引率

7.00%

发文量

1817

审稿时长

14 weeks

期刊介绍： Frontiers in Cellular and Infection Microbiology is a leading specialty journal, publishing rigorously peer-reviewed research across all pathogenic microorganisms and their interaction with their hosts. Chief Editor Yousef Abu Kwaik, University of Louisville is supported by an outstanding Editorial Board of international experts. This multidisciplinary open-access journal is at the forefront of disseminating and communicating scientific knowledge and impactful discoveries to researchers, academics, clinicians and the public worldwide. Frontiers in Cellular and Infection Microbiology includes research on bacteria, fungi, parasites, viruses, endosymbionts, prions and all microbial pathogens as well as the microbiota and its effect on health and disease in various hosts. The research approaches include molecular microbiology, cellular microbiology, gene regulation, proteomics, signal transduction, pathogenic evolution, genomics, structural biology, and virulence factors as well as model hosts. Areas of research to counteract infectious agents by the host include the host innate and adaptive immune responses as well as metabolic restrictions to various pathogenic microorganisms, vaccine design and development against various pathogenic microorganisms, and the mechanisms of antibiotic resistance and its countermeasures.