Clinical Microbiology and Infectious Diseases Department, Hospital General Universitario Gregorio Marañón, Universidad Complutense de Madrid, Madrid, Spain.
PLoS One. 2013 Apr 19;8(4):e61545. doi: 10.1371/journal.pone.0061545. Print 2013.
Methods based on real-time polymerase chain reaction (PCR) can speed up the diagnosis of invasive aspergillosis but are limited by a lack of standardization. We evaluated the commercially available MycAssay™ Aspergillus test for the diagnosis of invasive aspergillosis in patients without hematological cancer. We prospectively collected 322 lower respiratory tract samples (November 2009-January 2011) from 175 patients with lower respiratory tract infection and the following predisposing conditions: solid cancer (16.8%), cirrhosis (16.8%), corticosteroid therapy (71.7%), HIV infection (15.6%), chronic obstructive pulmonary disease (COPD, 52.6%), solid organ transplantation (kidney [1.2%], heart [3%], liver [4.6%]), or none (3.5%). Specimens were obtained when clinically indicated and analyzed in the microbiology laboratory. Aspergillus DNA was extracted and amplified by means of MycXtra® and MycAssay™ Aspergillus. Aspergillus spp. was isolated from 65 samples (31 patients). According to the European Organization for Research and Treatment of Cancer and Bulpa's criteria (for patients with COPD), 15 had probable invasive aspergillosis. MycAssay™ Aspergillus results were negative (n = 254), positive (n = 54), or indeterminate (n = 14). The sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic odds ratio of the MycAssay™ (first sample/any sample) were 86.7/93, 87.6/82.4, 34.1/34.1, 92.2/100, and 48/68.75. The differences between the proportion of samples with positive PCR determinations (63%) and the proportion of samples with Aspergillus spp. isolation (75%) did not reach statistical significance (P = 0.112). The median time from sample culture to visualization of fungal growth was 3 days, compared with ∼4 hours for MycAssay™ Aspergillus PCR. MycAssay™ Aspergillus showed high sensitivity for the diagnosis of invasive aspergillosis in patients without hematological cancer. Sensitivity increased when multiple samples were used. Compared with fungal culture, PCR significantly reduced the time to diagnosis.
方法基于实时聚合酶链反应 (PCR) 可以加速侵袭性曲霉病的诊断,但受到缺乏标准化的限制。我们评估了市售的 MycAssay™曲霉检测在没有血液恶性肿瘤的侵袭性曲霉病患者中的诊断价值。我们前瞻性收集了 175 例下呼吸道感染和以下易患疾病患者的 322 份下呼吸道样本(2009 年 11 月至 2011 年 1 月):实体瘤(16.8%)、肝硬化(16.8%)、皮质类固醇治疗(71.7%)、HIV 感染(15.6%)、慢性阻塞性肺疾病(COPD,52.6%)、实体器官移植(肾脏[1.2%]、心脏[3%]、肝脏[4.6%])或无(3.5%)。根据临床指征采集标本,并在微生物实验室进行分析。通过 MycXtra® 和 MycAssay™ Aspergillus 提取和扩增曲霉 DNA。从 65 份样本(31 例患者)中分离出曲霉属。根据欧洲癌症研究与治疗组织和 Bulpa 的标准(用于 COPD 患者),15 例为疑似侵袭性曲霉病。MycAssay™ Aspergillus 结果为阴性(n=254)、阳性(n=54)或不确定(n=14)。MycAssay™(首次样本/任何样本)的敏感性、特异性、阳性预测值、阴性预测值和诊断比值比分别为 86.7/93、87.6/82.4、34.1/34.1、92.2/100 和 48/68.75。PCR 检测阳性的样本比例(63%)和分离出曲霉属的样本比例(75%)之间的差异无统计学意义(P=0.112)。从样本培养到真菌生长可视化的中位数时间为 3 天,而 MycAssay™ Aspergillus PCR 约为 4 小时。MycAssay™ Aspergillus 对无血液恶性肿瘤的侵袭性曲霉病患者的诊断具有较高的敏感性。当使用多个样本时,敏感性增加。与真菌培养相比,PCR 显著缩短了诊断时间。