Rafanomezantsoa Lovanirina Clémencia, Sabourin Estelle, Guennouni Sebbouh Nadia, Sitterlé Emilie, Ben Halima Nada, Raveloarisaona Yannick Sonjah, Quesne Gilles, Dannaoui Eric, Bougnoux Marie-Elisabeth
Université Paris Cité, Faculté de Médecine, APHP, Hôpital Necker Enfants-Malades, Hôpital Européen Georges Pompidou, Unité de Parasitologie-Mycologie, Service de Microbiologie, Paris, France.
Dynamyc Research Group, Paris Est Créteil University (UPEC, EnvA), Paris, France.
Microbiol Spectr. 2024 Aug 6;12(8):e0358523. doi: 10.1128/spectrum.03585-23. Epub 2024 Jun 25.
Mucormycosis is a severe and emerging invasive fungal infection associated with high mortality rates. Early diagnosis is crucial for initiating specific antifungal treatment, with molecular tools currently representing the most efficient diagnostic approach. Presently, a standardized in-house real-time PCR method is widely employed for diagnosing mucormycosis. Our study aimed to evaluate the agreement for the Mucorales DNA detection between two commercial real-time PCR assays-the Fungiplex Mucorales Real-Time PCR Kit and the MycoGENIE spp. Real-Time PCR Kit-in comparison with the in-house PCR. We retrospectively analyzed 58 samples previously identified as positive for Mucorales using the in-house PCR. These samples, obtained from 22 patients with proven or probable mucormycosis, were tested with both commercial kits. Additionally, samples from 40 patients without mucormycosis served as negative controls. Our findings revealed that the MycoGENIE Kit demonstrated superior performance in detecting Mucorales DNA in samples identified as positive by the in-house PCR. Notably, we observed minimal variability in cycle threshold (CT) values when comparing the results of the MycoGENIE Kit with those of the in-house PCR, with an average difference of 1.8 cycles. In contrast, the Fungiplex Kit exhibited a larger discrepancy in CT values compared to the in-house PCR, with an average difference of 4.1 cycles. The MycoGENIE Kit exhibited very good agreement (kappa of 0.82) with the in-house PCR for detecting Mucorales DNA across various sample types. These findings are important for the choice of kits that could be used to diagnose mucormycosis in clinical microbiology laboratories.
Early diagnosis of mucormycosis is crucial for initiating effective treatment. The detection of Mucorales DNA by PCR in serum has revolutionized the diagnosis of this infection. However, the use of in-house methods can be time consuming. The availability of a commercial kit eliminates the need for in-house assay development, reducing laboratory workload and ensuring consistent performance across different healthcare settings. Currently, there are several commercial assays available, but many have limited evaluation. In this study, we compared two commercial kits and found that the MycoGENIE Kit offers a promising alternative to the in-house method.
毛霉病是一种严重且新出现的侵袭性真菌感染，死亡率很高。早期诊断对于启动特定的抗真菌治疗至关重要，分子工具目前是最有效的诊断方法。目前，一种标准化的内部实时PCR方法被广泛用于诊断毛霉病。我们的研究旨在评估两种商业实时PCR检测试剂盒（Fungiplex毛霉目实时PCR试剂盒和MycoGENIE spp.实时PCR试剂盒）与内部PCR相比，在检测毛霉目DNA方面的一致性。我们回顾性分析了58份先前使用内部PCR鉴定为毛霉目阳性的样本。这些样本来自22例确诊或可能患有毛霉病的患者，用两种商业试剂盒进行检测。此外，来自40例无毛霉病患者的样本用作阴性对照。我们的研究结果表明，MycoGENIE试剂盒在检测内部PCR鉴定为阳性的样本中的毛霉目DNA方面表现出更好的性能。值得注意的是，将MycoGENIE试剂盒的结果与内部PCR的结果进行比较时，我们观察到循环阈值（CT）值的变异性最小，平均差异为1.8个循环。相比之下，Fungiplex试剂盒与内部PCR相比，CT值差异更大，平均差异为4.1个循环。MycoGENIE试剂盒在检测各种样本类型中的毛霉目DNA方面与内部PCR表现出非常好的一致性（kappa值为0.82）。这些发现对于临床微生物实验室中可用于诊断毛霉病的试剂盒的选择很重要。
毛霉病的早期诊断对于启动有效治疗至关重要。通过PCR检测血清中的毛霉目DNA彻底改变了这种感染的诊断。然而，使用内部方法可能很耗时。商业试剂盒的可用性消除了开发内部检测方法的需要，减少了实验室工作量，并确保了不同医疗环境下的一致性能。目前有几种商业检测方法可用，但许多评估有限。在本研究中，我们比较了两种商业试剂盒，发现MycoGENIE试剂盒为内部方法提供了一种有前景的替代方案。
