背景：这项研究评估了一种新型的快速宽范围PCR和测序（FBR-PCR/S）测定法的性能，以改善加拿大大型医疗保健地区高危患者的侵袭性真菌病（IFD）的诊断。
方法：在2年的时间内，对107例患者的114份临床标本（CS），包括支气管肺泡灌洗（BAL）进行了前瞻性检测。还测试了接种了已知真菌病原体的BAL（n=33）以增加多样性。患者特征,从实验室信息系统收集真菌染色和培养结果。使用靶向内部转录间隔区（ITS）（〜350bp）和大亚基（LSU）（〜550bp）基因区域的双引发寡核苷酸（DPO）引物对提取的BALs进行FBR-PCR/S测定/CS。对照标准微生物学方法和关于IFD存在的临床评价来评估分子测试的性能。
结果：107例患者主要为男性（67，62.6％），平均年龄为59岁（范围=0-85岁）：74例（69.2％）患者至少有一种潜在的合并症：19例（34.5％）已确诊，12例（21.8％）有可能的IFD。培养物从55个念珠菌属BALs/CS中回收了66个真菌分离株。和曲霉属。是最常见的。对于BAL,分子测定与标准方法具有灵敏度，特异性，阳性预测值(PPV)和阴性预测值(NPV)，效率为88.5%vs.100%，100%vs.61.1%,100%vs.88.5%,61.1%vs.100%,和90.2%。对于其他CS，分子测定具有与标准方法相似的性能，具有灵敏度，特异性，PPV,净现值和效率为66.7%，87.0%,66.7%,两种方法均为87.0%和81.3%。这两种方法也类似地执行，无论CS染色/显微镜是否显示酵母/真菌元素。与花费4至6周的真菌培养物相比,FBR-PCR/S测定结果在〜8小时内报告。
结论：与标准方法相比，快速分子检测具有同等的诊断效率，但通过在相同的日移（约8小时）报告快速物种水平鉴定，提高了临床实用性。
BACKGROUND: This study evaluated the performance of a novel fast broad range PCR and sequencing (FBR-PCR/S) assay for the improved diagnosis of invasive fungal disease (IFD) in high-risk patients in a large Canadian healthcare region.
METHODS: A total of 114 clinical specimens (CS) including bronchoalveolar lavages (BALs) were prospectively tested from 107 patients over a 2-year period. Contrived BALs (n = 33) inoculated with known fungi pathogens were also tested to increase diversity. Patient characteristics, fungal stain and culture results were collected from the laboratory information system. Dual-priming oligonucleotide (DPO) primers targeted to the internal transcribed spacer (ITS) (~ 350 bp) and large subunit (LSU) (~ 550 bp) gene regions were used to perform FBR-PCR/S assays on extracted BALs/CS. The performance of the molecular test was evaluated against standard microbiological methods and clinical review for the presence of IFD.
RESULTS: The 107 patients were predominantly male (67, 62.6%) with a mean age of 59 years (range = 0-85 years): 74 (69.2%) patients had at least one underlying comorbidity: 19 (34.5%) had confirmed and 12 (21.8%) had probable IFD. Culture recovered 66 fungal isolates from 55 BALs/CS with Candida spp. and Aspergillus spp. being most common. For BALs, the molecular assay vs. standard methods had sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV), and efficiency of 88.5% vs.100%, 100% vs. 61.1%, 100% vs. 88.5%, 61.1% vs. 100%, and 90.2% for both. For other CS, the molecular assay had similar performance to standard methods with sensitivity, specificity, PPV, NPV and efficiency of 66.7%, 87.0%, 66.7%, 87.0% and 81.3% for both methods. Both methods also performed similarly, regardless of whether CS stain/microscopy showed yeast/fungal elements. FBR-PCR/S assays results were reported in ~ 8 h compared to fungal cultures that took between 4 and 6 weeks.
CONCLUSIONS: Rapid molecular testing compared to standard methods have equivalent diagnostic efficiency but improves clinical utility by reporting a rapid species-level identification the same dayshift (~ 8 h).
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