Mah Jordan, Lieu Anthony, Nicholas Veronica, Moreno Angel, Murugesan Kanagavel, Budvytiene Indre, Banaei Niaz
Department of Pathology, Stanford University School of Medicine, Stanford, California, USA.
Clinical Microbiology Laboratory, Stanford Health Care, Stanford, California, USA.
J Clin Microbiol. 2025 Aug 13;63(8):e0079625. doi: 10.1128/jcm.00796-25. Epub 2025 Jul 9.
Diagnosis of mucormycosis poses a substantial challenge due to the lack of a non-invasive biomarker and limitations of conventional diagnostics using invasive specimens. The aim of this study was to characterize the performance of Mucorales plasma cell-free DNA (cfDNA) PCR in immunosuppressed and non-immunosuppressed patients with pulmonary, disseminated, and localized mucormycosis. Patients with Mucorales plasma cfDNA test results were retrospectively categorized as proven or probable for sensitivity analysis and as no Mucorales infection for specificity analysis. In total, 85 positive and 212 negative Mucorales plasma cfDNA test results in unique patients were included in this study. Per the expert definitions, 47 patients had proven or probable mucormycosis, and 171 did not have invasive Mucorales infection. The Mucorales plasma cfDNA PCR had an overall sensitivity and specificity of 85.1% (40/47, 95% CI, 71.7-93.8) and 92.9% (158/170, 95% CI, 88.0-96.3), respectively. The sensitivity was 92.1% (35/38, 95% CI, 78.6-98.3) and 55.6% (5/9, 95% CI, 21.2-86.3) in patients with and without immunosuppression, respectively, and 100% (14/14, 95% CI, 76.8-100), 89.5% (17/19, 95% CI, 66.9-98.7), and 64.3% (9/14, 95% CI, 35.1-87.2) in patients with disseminated, pulmonary, and localized mucormycosis, respectively. Mucorales plasma cfDNA PCR is a sensitive and specific non-invasive testing modality for the diagnosis of mucormycosis in immunosuppressed patients and those with pulmonary and disseminated infection.IMPORTANCEMucormycosis is an invasive mold infection associated with high morbidity and mortality. Early diagnosis and effective antifungal treatment are critical for improving clinical outcomes. However, diagnosis of mucormycosis is often delayed due to the lack of a non-invasive biomarker and insensitivity of culture and non-specificity of histopathology performed on invasive specimens. Mucorales plasma cell-free DNA (cfDNA) PCR is a novel testing modality that allows non-invasive diagnosis of mucormycosis. In the current study, we evaluated the clinical performance of a pre-analytically optimized Mucorales plasma cfDNA PCR in immunosuppressed and non-immunosuppressed patients with pulmonary, disseminated, and localized infections. We show that Mucorales plasma cfDNA PCR is highly sensitive and specific in immunosuppressed patients with pulmonary and disseminated infections. Thus, the Mucorales plasma cfDNA PCR represents an accurate diagnostic tool for non-invasive diagnosis of mucormycosis, which may enable early treatment and improved outcomes in immunosuppressed patients with mucormycosis.
由于缺乏非侵入性生物标志物以及使用侵入性标本的传统诊断方法存在局限性，毛霉菌病的诊断面临重大挑战。本研究的目的是评估毛霉目无细胞血浆DNA（cfDNA）PCR在免疫抑制和非免疫抑制的肺部、播散性和局限性毛霉菌病患者中的诊断性能。对有毛霉目血浆cfDNA检测结果的患者进行回顾性分类，分为确诊或疑似用于敏感性分析，无毛霉目感染用于特异性分析。本研究共纳入了85例毛霉目血浆cfDNA检测结果为阳性的患者和212例检测结果为阴性的患者。根据专家定义，47例患者确诊或疑似患有毛霉菌病，171例患者没有侵袭性毛霉目感染。毛霉目血浆cfDNA PCR的总体敏感性和特异性分别为85.1%（40/47，95%CI，71.7-93.8）和92.9%（158/170，95%CI，88.0-96.3）。免疫抑制患者和非免疫抑制患者的敏感性分别为92.1%（35/38，95%CI，78.6-98.3）和55.6%（5/9，95%CI，21.2-86.3），播散性、肺部和局限性毛霉菌病患者的敏感性分别为100%（14/14，95%CI，76.8-100）、89.5%（17/19，95%CI，66.9-98.7）和64.3%（9/14，95%CI，35.1-87.2）。毛霉目血浆cfDNA PCR是一种敏感且特异的非侵入性检测方法，可用于诊断免疫抑制患者以及患有肺部和播散性感染的患者的毛霉菌病。
重要性
毛霉菌病是一种侵袭性霉菌感染，与高发病率和死亡率相关。早期诊断和有效的抗真菌治疗对于改善临床结局至关重要。然而，由于缺乏非侵入性生物标志物以及培养的不敏感性和对侵入性标本进行组织病理学检查的非特异性，毛霉菌病的诊断常常延迟。毛霉目无细胞血浆DNA（cfDNA）PCR是一种新型检测方法，可实现毛霉菌病的非侵入性诊断。在本研究中，我们评估了一种经过分析前优化的毛霉目血浆cfDNA PCR在免疫抑制和非免疫抑制的肺部、播散性和局限性感染患者中的临床性能。我们发现，毛霉目血浆cfDNA PCR在免疫抑制的肺部和播散性感染患者中具有高度敏感性和特异性。因此，毛霉目血浆cfDNA PCR是一种用于毛霉菌病非侵入性诊断的确切诊断工具，这可能使免疫抑制的毛霉菌病患者能够早期治疗并改善结局。
