反转录酶实时荧光定量PCR技术在曲霉菌侵袭性感染诊断策略中的应用。

IF 6.1 2区 医学 Q1 MICROBIOLOGY
Journal of Clinical Microbiology Pub Date : 2024-11-13 Epub Date: 2024-10-24 DOI:10.1128/jcm.00791-24
Charles Gibert, Pauline Tirard-Collet, Charline Miossec, Damien Dupont, Florence Persat, Martine Wallon, Florence Ader, Gilles Devouassoux, Sophie Ducastelle, Hélène Labussière-Wallet, Sylvie Paulus, Céline Guichon, Anne-Claire Lukaszewicz, Jean-Christophe Richard, Florent Wallet, Alexandre Alanio, Meja Rabodonirina, Jean Menotti
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引用次数: 0

摘要

我们的目的是开发一种针对曲霉菌的 RT-qPCR,并比较其与烟曲霉菌 qPCR 在诊断侵袭性曲霉菌病(IA)方面的性能。里昂大学医院在两年内对怀疑患有侵袭性曲霉菌病的患者进行了曲霉菌 qPCR 分子诊断检测。根据欧洲癌症研究和治疗组织/霉菌病研究小组(EORTC-MSGERC)的疑似 IA 标准对患者进行了分类;对所有纳入的样本进行了 RT-qPCR 和 qPCR 检测。以 EORTC-MSGERC 分类结果为参考,计算并比较了 RT-qPCR 和 qPCR 的敏感性和特异性。根据 IA 分类和样本类型对周期阈值(Ct)结果进行了比较。在分析的 193 份样本中,91 份被归类为排除 IA,46 份被归类为可能 IA,53 份被归类为可能 IA,3 份被归类为已证实 IA。在所有样本类型中,RT-qPCR 对所有 IA 分类的灵敏度明显高于 qPCR,多检测出 17/102 个样本(P 值 < 0.01)。对于血浆样本,使用 RT-qPCR 检测所有 IA 分类的灵敏度明显更高,特异性明显更低(P 值 < 0.001)。就所有 IA 分类和所有样本类型而言,使用 RT-qPCR 获得的平均 Ct 值明显低于使用 qPCR 获得的平均 Ct 值(P 值分别 < 0.001 和 P 值 < 0.0001)。在诊断由烟曲霉引起的侵袭性曲霉病时,RT-qPCR 比 qPCR 具有更高的灵敏度,尤其是在真菌含量固有较低的样本中。重要意义烟曲霉属于世界卫生组织真菌优先病原体名单中的关键优先组。侵袭性曲霉菌病(IA)是一种危及生命的感染,预后不良,诊断困难。PCR 已被纳入 2020 年欧洲癌症研究和治疗组织/真菌病研究小组关于 IA 诊断的共识定义。然而,由于真菌负担经常较低,其灵敏度有待提高。本研究提出了一种检测核糖体RNA和DNA总核酸的创新方法,与传统技术相比,该方法可提高IA诊断的灵敏度,尤其是在血液等非侵入性样本中,从而加强对高危患者感染情况的监测。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Reverse-transcriptase real-time PCR in the diagnostic strategy for invasive infections caused by Aspergillus fumigatus.

The aim was to develop an RT-qPCR targeting Aspergillus fumigatus and compare its performance to that of Aspergillus fumigatus qPCR for the diagnosis of invasive aspergillosis (IA). Samples from patients of the Lyon University hospitals for whom a suspicion of IA led to the realization of an Aspergillus fumigatus qPCR molecular diagnostic test over a 2-year period were included. The patients were classified according to the European Organization for Research and Treatment of Cancer/Mycoses Study Group (EORTC-MSGERC) criteria for suspected IA; RT-qPCR and qPCR assays were performed on all included samples. The sensitivities and specificities of RT-qPCR and qPCR were calculated and compared using the results of the EORTC-MSGERC classification as reference. The cycle threshold (Ct) results were compared according to IA classification and sample type. Among the 193 samples analyzed, 91 were classified as IA excluded, 46 as possible IA, 53 as probable IA, and 3 as proven IA. For all sample types, RT-qPCR was significantly more sensitive than qPCR for all IA classifications with an additional 17/102 samples detected (P-value < 0.01). For plasma samples, sensitivity was significantly higher and specificity significantly lower using RT-qPCR for all IA classifications (P-value < 0.001). The mean Ct obtained with RT-qPCR were significantly lower than those obtained with qPCR for all IA classifications and all sample types (P-value < 0.001 and P-value < 0.0001, respectively). RT-qPCR presents a higher sensitivity than qPCR for the diagnosis of IA due to Aspergillus fumigatus, particularly in samples with an intrinsically low fungal load.IMPORTANCEAspergillus fumigatus belongs to the critical priority group of the World Health Organization fungal priority pathogens list. Invasive aspergillosis (IA) is a life-threatening infection with poor prognosis and challenging diagnosis. PCR has been integrated into the 2020 European Organization for Research and Treatment of Cancer/Mycoses Study Group consensus definitions for IA diagnosis. However, due to frequent low fungal burdens, its sensitivity needs to be improved. This work presents an innovative method for detecting total nucleic acids, corresponding to both ribosomal RNA and DNA, that enables IA diagnosis with greater sensitivity than conventional techniques, especially in non-invasive samples such as blood, enhancing the monitoring of this infection in high-risk patients.

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来源期刊
Journal of Clinical Microbiology
Journal of Clinical Microbiology 医学-微生物学
CiteScore
17.10
自引率
4.30%
发文量
347
审稿时长
3 months
期刊介绍: The Journal of Clinical Microbiology® disseminates the latest research concerning the laboratory diagnosis of human and animal infections, along with the laboratory's role in epidemiology and the management of infectious diseases.
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