Brown Lottie, Cruciani Mario, Morton Charles Oliver, Alanio Alexandre, Barnes Rosemary A, Donnelly J Peter, Hagen Ferry, Gorton Rebecca, Lackner Michaela, Loeffler Juergen, Millon Laurence, Rautemaa-Richardson Riina, White P Lewis
Institute of Infection and Immunity, St. George's University and St. Georges University Hospitals NHS Foundation Trust, London SW17 0QT, UK.
Fungal PCR Initiative, A Working Group of the International Society of Human and Animal Mycology, 37100 Verona, Italy.
Diagnostics (Basel). 2025 Jul 30;15(15):1909. doi: 10.3390/diagnostics15151909.
: Polymerase chain reaction (PCR) is highly sensitive and specific for the rapid diagnosis of invasive fungal disease (IFD) but is not yet widely implemented due to concerns regarding limited standardisation between assays, the lack of commercial options and the absence of clear guidance on interpreting results. : This review provides an update on technical and clinical aspects of PCR for the diagnosis of the most pertinent fungal pathogens, including , , , spp., and endemic mycoses. : Recent meta-analyses have demonstrated that quantitative PCR (qPCR) offers high sensitivity for diagnosing IFD, surpassing conventional microscopy, culture and most serological tests. The reported specificity of qPCR is likely underestimated due to comparison with imperfect reference standards with variable sensitivity. Although the very low limit of detection of qPCR can generate false positive results due to procedural contamination or patient colonisation (particularly in pulmonary specimens), the rates are comparable to those observed for biomarker testing. When interpreting qPCR results, it is essential to consider the pre-test probability, determined by the patient population, host factors, clinical presentation and risk factors. For patients with low to moderate pre-test probability, the use of sensitive molecular tests, often in conjunction with serological testing or biomarkers, can effectively exclude IFD when all tests return negative results, reducing the need for empirical antifungal therapy. Conversely, for patients with high pre-test probability and clinical features of IFD, qPCR testing on invasive specimens from the site of infection (such as tissue or bronchoalveolar lavage fluid) can confidently rule in the disease. The development of next-generation sequencing methods to detect fungal infection has the potential to enhance the diagnosis of IFD, but standardisation and optimisation are essential, with improved accessibility underpinning clinical utility.
聚合酶链反应（PCR）对侵袭性真菌病（IFD）的快速诊断具有高度敏感性和特异性，但由于检测方法之间标准化有限、缺乏商业选择以及缺乏结果解读的明确指南等问题，尚未得到广泛应用。
本综述提供了关于PCR在诊断最相关真菌病原体方面的技术和临床方面的最新信息，包括曲霉属、念珠菌属、隐球菌属、接合菌纲真菌以及地方性真菌病。
最近的荟萃分析表明，定量PCR（qPCR）对IFD的诊断具有高敏感性，超过了传统显微镜检查、培养和大多数血清学检测。由于与敏感性可变的不完善参考标准进行比较，qPCR报道的特异性可能被低估。尽管qPCR极低的检测限可能因操作污染或患者定植（特别是在肺部标本中）而产生假阳性结果，但其发生率与生物标志物检测中观察到的相当。在解读qPCR结果时，必须考虑由患者群体、宿主因素、临床表现和危险因素确定的检测前概率。对于检测前概率低至中等的患者，使用敏感的分子检测，通常与血清学检测或生物标志物联合使用，当所有检测结果均为阴性时，可有效排除IFD，减少经验性抗真菌治疗的需求。相反，对于检测前概率高且具有IFD临床特征的患者，对来自感染部位（如组织或支气管肺泡灌洗液）的侵袭性标本进行qPCR检测可明确诊断该病。检测真菌感染的下一代测序方法的发展有可能提高IFD的诊断水平，但标准化和优化至关重要，提高可及性是临床应用的基础。
