Pandey Mragnayani, Xess Immaculata, Sachdev Janya, Yadav Usha, Singh Gagandeep, Pradhan Dibyabhaba, Xess Ashit Bhushan, Rana Bhaskar, Dar Lalit, Bakhshi Sameer, Seth Rachna, Mahapatra Manoranjan, Jyotsna Viveka P, Jain Arun Kumar, Kumar Rakesh, Agarwal Reshu, Mani Prashant
Department of Microbiology, All India Institute of Medical Sciences, New Delhi, India.
ICMR Computational Genomics Centre Informatics, Systems & Research Management Division Indian Council of Medical Research, All India Institute of Medical Sciences, New Delhi, India.
Front Fungal Biol. 2022 Jan 24;2:800898. doi: 10.3389/ffunb.2021.800898. eCollection 2021.
Molecular diagnostic assays can expedite the diagnosis of fungal infections, and subsequently help in early interventions and appropriate management of patients. The aim of this study was to develop a single set of primers for a real-time quantitative polymerase chain reaction (qPCR) assay to detect and identify commonly reported, clinically relevant molds i.e., and , up to genus level by melting curve analysis. This assay was evaluated in whole blood from patients with suspected invasive aspergillosis (IA), and in tissue biopsy, bronchoalveolar lavage (BAL) fluid and other site-specific samples from patients with suspected invasive mucormycosis (IM). The limit of detection (LoD) was determined as 10 copies/μl for all three molds. The mean coefficient of variation (CV) across all sets of intra- and inter-assay data was 0.63% (ranging from 0.42 to 1.56%), showing high reproducibility of the assay. Sensitivity and specificity of the assay were 93.3 and 97.1% respectively for diagnosis of IA, and 99.29 and 83.84% respectively for diagnosis of IM. was not detected in any of the clinical samples included and the few laboratory confirmed cases of fusariosis did not meet the inclusion criteria of the study. Hence no ROC curve or cutoff value could be generated for the same. This newly developed qPCR assay therefore appears to be a promising tool in detection of IA and IM.
分子诊断检测可加快真菌感染的诊断，并随后有助于对患者进行早期干预和适当管理。本研究的目的是开发一套用于实时定量聚合酶链反应（qPCR）检测的引物，通过熔解曲线分析在属水平上检测和鉴定常见的、临床相关的霉菌，即 和 。该检测方法在疑似侵袭性曲霉病（IA）患者的全血中，以及在疑似侵袭性毛霉病（IM）患者的组织活检、支气管肺泡灌洗（BAL）液和其他特定部位样本中进行了评估。所有三种霉菌的检测限（LoD）均确定为10拷贝/μl。所有组内和组间检测数据的平均变异系数（CV）为0.63%（范围为0.42%至1.56%），表明该检测方法具有高重复性。该检测方法诊断IA的敏感性和特异性分别为93.3%和97.1%，诊断IM的敏感性和特异性分别为99.29%和83.84%。在所纳入的任何临床样本中均未检测到 ，少数实验室确诊的镰刀菌病病例不符合该研究的纳入标准。因此，无法为其生成ROC曲线或临界值。因此，这种新开发的qPCR检测方法似乎是检测IA和IM的一种有前景的工具。
