Hansheng Wang, Xiao Chen, Xiaofeng Wu, Qizhen Cao, Yi Wu, Fang Wang, Yunyun Wang, Yanhui Zhou, Yijun Tang, Tao Ren, Meifang Wang
{"title":"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","authors":"Hansheng Wang, Xiao Chen, Xiaofeng Wu, Qizhen Cao, Yi Wu, Fang Wang, Yunyun Wang, Yanhui Zhou, Yijun Tang, Tao Ren, Meifang Wang","doi":"10.3389/fcimb.2025.1611391","DOIUrl":null,"url":null,"abstract":"<p><strong>Background and objective: </strong><i>Pneumocystis jirovecii</i> pneumonia (PjP) remains an important cause of morbimortality worldwide, and differentiating <i>Pneumocystis jirovecii</i> (<i>P. jirovecii</i>) infection from <i>P. jirovecii</i> 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 <i>P. jirovecii</i> identification in bronchoalveolar lavage fluid (BALF) samples combined with serum 1,3-β-D-glucan (BDG) level detection to differentiate PjP and PjC.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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 (<i>p <</i>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 (<i>p <</i>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%).</p><p><strong>Conclusion: </strong>BALF mp-tNGS and serum BDG serve as valuable adjunct diagnostic tools, providing reliable differentiation between <i>P. jirovecii</i> colonization and active infection.</p>","PeriodicalId":12458,"journal":{"name":"Frontiers in Cellular and Infection Microbiology","volume":"15 ","pages":"1611391"},"PeriodicalIF":4.8000,"publicationDate":"2025-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12489942/pdf/","citationCount":"0","resultStr":"{\"title\":\"Combined multiplex polymerase chain reaction-based targeted next-generation sequencing and serum 1, 3-β-D-glucan for differential diagnosis of <i>Pneumocystis</i> pneumonia <i>and Pneumocystis</i> colonization.\",\"authors\":\"Hansheng Wang, Xiao Chen, Xiaofeng Wu, Qizhen Cao, Yi Wu, Fang Wang, Yunyun Wang, Yanhui Zhou, Yijun Tang, Tao Ren, Meifang Wang\",\"doi\":\"10.3389/fcimb.2025.1611391\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background and objective: </strong><i>Pneumocystis jirovecii</i> pneumonia (PjP) remains an important cause of morbimortality worldwide, and differentiating <i>Pneumocystis jirovecii</i> (<i>P. jirovecii</i>) infection from <i>P. jirovecii</i> 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 <i>P. jirovecii</i> identification in bronchoalveolar lavage fluid (BALF) samples combined with serum 1,3-β-D-glucan (BDG) level detection to differentiate PjP and PjC.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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 (<i>p <</i>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 (<i>p <</i>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%).</p><p><strong>Conclusion: </strong>BALF mp-tNGS and serum BDG serve as valuable adjunct diagnostic tools, providing reliable differentiation between <i>P. jirovecii</i> colonization and active infection.</p>\",\"PeriodicalId\":12458,\"journal\":{\"name\":\"Frontiers in Cellular and Infection Microbiology\",\"volume\":\"15 \",\"pages\":\"1611391\"},\"PeriodicalIF\":4.8000,\"publicationDate\":\"2025-09-18\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12489942/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Frontiers in Cellular and Infection Microbiology\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.3389/fcimb.2025.1611391\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2025/1/1 0:00:00\",\"PubModel\":\"eCollection\",\"JCR\":\"Q2\",\"JCRName\":\"IMMUNOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Frontiers in Cellular and Infection Microbiology","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.3389/fcimb.2025.1611391","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/1/1 0:00:00","PubModel":"eCollection","JCR":"Q2","JCRName":"IMMUNOLOGY","Score":null,"Total":0}
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.
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.
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.
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