Morton C Oliver, White P Lewis, Barnes Rosemary A, Klingspor Lena, Cuenca-Estrella Manuel, Lagrou Katrien, Bretagne Stéphane, Melchers Willem, Mengoli Carlo, Caliendo Angela M, Cogliati Massimo, Debets-Ossenkopp Yvette, Gorton Rebecca, Hagen Ferry, Halliday Catriona, Hamal Petr, Harvey-Wood Kathleen, Jaton Katia, Johnson Gemma, Kidd Sarah, Lengerova Martina, Lass-Florl Cornelia, Linton Chris, Millon Laurence, Morrissey C Orla, Paholcsek Melinda, Talento Alida Fe, Ruhnke Markus, Willinger Birgit, Donnelly J Peter, Loeffler Juergen
Western Sydney University, Sydney, Australia.
Public Health Wales Microbiology Cardiff, UK.
Med Mycol. 2017 Jun 1;55(4):402-413. doi: 10.1093/mmy/myw093.
A wide array of PCR tests has been developed to aid the diagnosis of invasive aspergillosis (IA), providing technical diversity but limiting standardisation and acceptance. Methodological recommendations for testing blood samples using PCR exist, based on achieving optimal assay sensitivity to help exclude IA. Conversely, when testing more invasive samples (BAL, biopsy, CSF) emphasis is placed on confirming disease, so analytical specificity is paramount. This multicenter study examined the analytical specificity of PCR methods for detecting IA by blind testing a panel of DNA extracted from a various fungal species to explore the range of Aspergillus species that could be detected, but also potential cross reactivity with other fungal species. Positivity rates were calculated and regression analysis was performed to determine any associations between technical specifications and performance. The accuracy of Aspergillus genus specific assays was 71.8%, significantly greater (P < .0001) than assays specific for individual Aspergillus species (47.2%). For genus specific assays the most often missed species were A. lentulus (25.0%), A. versicolor (24.1%), A. terreus (16.1%), A. flavus (15.2%), A. niger (13.4%), and A. fumigatus (6.2%). There was a significant positive association between accuracy and using an Aspergillus genus PCR assay targeting the rRNA genes (P = .0011). Conversely, there was a significant association between rRNA PCR targets and false positivity (P = .0032). To conclude current Aspergillus PCR assays are better suited for detecting A. fumigatus, with inferior detection of most other Aspergillus species. The use of an Aspergillus genus specific PCR assay targeting the rRNA genes is preferential.
已经开发出一系列PCR检测方法来辅助侵袭性曲霉病（IA）的诊断，这提供了技术多样性，但限制了标准化和可接受性。基于实现最佳检测灵敏度以帮助排除IA，存在关于使用PCR检测血样的方法学建议。相反，在检测更具侵入性的样本（支气管肺泡灌洗、活检、脑脊液）时，重点在于确诊疾病，因此分析特异性至关重要。这项多中心研究通过对从各种真菌物种中提取的一组DNA进行盲测，检查了用于检测IA的PCR方法的分析特异性，以探索可检测的曲霉物种范围，以及与其他真菌物种的潜在交叉反应性。计算阳性率并进行回归分析，以确定技术规格与性能之间的任何关联。曲霉属特异性检测的准确率为71.8%，显著高于单个曲霉物种特异性检测（47.2%）（P < .0001）。对于属特异性检测，最常漏检的物种是勒图曲霉（25.0%）、杂色曲霉（24.1%）、土曲霉（16.1%）、黄曲霉（15.2%）、黑曲霉（13.4%）和烟曲霉（6.2%）。准确率与使用靶向rRNA基因的曲霉属PCR检测之间存在显著正相关（P = .0011）。相反，rRNA PCR靶点与假阳性之间存在显著关联（P = .0032）。总之，目前的曲霉PCR检测更适合检测烟曲霉，对大多数其他曲霉物种的检测效果较差。优先使用靶向rRNA基因的曲霉属特异性PCR检测。
