血浆无细胞游离 DNA 真菌聚合酶链反应检测 panel 对非侵袭性真菌感染诊断的临床准确性和影响。

Clinical Accuracy and Impact of Plasma Cell-Free DNA Fungal Polymerase Chain Reaction Panel for Noninvasive Diagnosis of Fungal Infection.

机构信息

Department of Pathology, Stanford University School of Medicine, Stanford, California, USA.

Clinical Microbiology Laboratory, Stanford University Medical Center, Palo Alto, California, USA.

出版信息

Clin Infect Dis. 2021 Nov 2;73(9):1677-1684. doi: 10.1093/cid/ciab158.

DOI:10.1093/cid/ciab158

PMID:33606010

Abstract

BACKGROUND

Invasive fungal infection (IFI) is a growing cause of morbidity and mortality in oncology and transplant patients. Diagnosis of IFI is often delayed due to need for invasive biopsy and low sensitivity of conventional diagnostic methods. Fungal cell-free DNA (cfDNA) detection in plasma is a novel testing modality for the noninvasive diagnosis of IFI.

METHODS

A novel bioinformatic pipeline was created to interrogate fungal genomes and identify multicopy sequences for cfDNA polymerase chain reaction (PCR) targeting. A real-time PCR panel was developed for 12 genera and species most commonly causing IFI. Sensitivity and specificity of the fungal PCR panel were determined using plasma samples from patients with IFI and non-IFI controls. Clinical impact of the fungal PCR panel was evaluated prospectively based on the treating team's interpretation of the results.

RESULTS

Overall, the sensitivity and specificity were 56.5% (65/115; 95% confidence interval [CI], 47.4-65.2) and 99.5% (2064/2075; 95% CI, 99.0-99.7), respectively. In the subset of patients with an optimized plasma volume (2 mL), sensitivity was 69.6% (48/69; 95% CI, 57.9-79.2). Sensitivity was 91.7% (11/12; 95% CI, 62.5-100) for detection of Mucorales agents, 56.3% (9/16; 95% CI, 33.2-76.9) for Aspergillus species, and 84.6% (11/13; 95% CI, 56.5-96.9) for Candida albicans. In a prospective evaluation of 226 patients with suspected IFI, cfDNA testing was positive in 47 (20.8%) patients and resulted in a positive impact on clinical management in 20 of 47 (42.6%).

CONCLUSIONS

The fungal cfDNA PCR panel offers a noninvasive approach to early diagnosis of IFI, providing actionable results for personalized care.

摘要

背景

侵袭性真菌感染（IFI）是肿瘤和移植患者发病率和死亡率不断上升的原因。由于需要进行侵入性活检和常规诊断方法的敏感性低，IFI 的诊断常常被延误。血浆中真菌无细胞 DNA（cfDNA）检测是一种用于IFI 非侵入性诊断的新型检测方法。

方法

创建了一种新的生物信息学管道，用于询问真菌基因组并识别用于 cfDNA 聚合酶链反应（PCR）靶向的多拷贝序列。开发了用于最常引起 IFI 的 12 个属和种的实时 PCR 检测试剂盒。使用 IFI 和非 IFI 对照组患者的血浆样本确定真菌 PCR 检测试剂盒的敏感性和特异性。根据治疗团队对结果的解释，前瞻性评估真菌 PCR 检测试剂盒的临床影响。

结果

总体而言，敏感性和特异性分别为 56.5%（65/115；95%置信区间[CI]，47.4-65.2）和 99.5%（2075/2075；95%CI，99.0-99.7）。在优化血浆量（2 mL）的患者亚组中，敏感性为 69.6%（48/69；95%CI，57.9-79.2）。检测毛霉目真菌的敏感性为 91.7%（11/12；95%CI，62.5-100），检测曲霉属的敏感性为 56.3%（9/16；95%CI，33.2-76.9），检测白色念珠菌的敏感性为 84.6%（11/13；95%CI，56.5-96.9）。在对 226 例疑似 IFI 患者的前瞻性评估中，cfDNA 检测在 47 例（20.8%）患者中呈阳性，在这 47 例患者中有 20 例（42.6%）对临床管理产生了积极影响。