Performance Evaluation of Five Real-Time PCR Assays for the Detection of Candida auris DNA.

IF 4.1 2区医学 Q1 DERMATOLOGY

Mycoses Pub Date : 2025-05-01 DOI:10.1111/myc.70065

Jochem B Buil, Bart van den Bosch, Suzan J van der Maas, Eelco F J Meijer, Theun de Groot, Joseph Meletiadis, Paul E Verweij, Willem J G Melchers, Suzan D Pas

{"title":"Performance Evaluation of Five Real-Time PCR Assays for the Detection of Candida auris DNA.","authors":"Jochem B Buil, Bart van den Bosch, Suzan J van der Maas, Eelco F J Meijer, Theun de Groot, Joseph Meletiadis, Paul E Verweij, Willem J G Melchers, Suzan D Pas","doi":"10.1111/myc.70065","DOIUrl":null,"url":null,"abstract":"Objectives: This study aimed to systematically evaluate and compare the performance of two laboratory-developed assays (LDAs) and three commercially available real-time PCR assays for the detection of Candida auris. The analytical sensitivity, specificity and limit of detection (LOD) of each assay were assessed, alongside their clinical sensitivity in identifying C. auris colonisation.Methods: Ten C. auris strains representing five clades, as well as genetically related yeasts, common yeast species, and dermatophytes, were used to assess assay sensitivity and cross reactivity. Clinical and environmental samples were collected from patients during an outbreak and tested with three commercial PCR assays (AurisID, Fungiplex, FungiXpert) and two LDAs (CDC LDA, EMC LDA). LOD was determined using Probit analysis. Diagnostic sensitivity was evaluated by comparing the detection rate of each individual assay to the total detection rate of all assays combined.Results: The EMC LDA exhibited the highest analytical sensitivity, with a LOD of 8 conidia/reaction, followed by CDC LDA (16 conidia/reaction), AurisID and FungiXpert (19 conidia/reaction), and Fungiplex (596 conidia/reaction). Specificity testing revealed cross-reactivity in the CDC LDA and AurisID assays with C. pseudohaemulonii at high conidia levels, while no cross-reactivity was observed in the other assays. EMC LDA showed the highest clinical sensitivity (100%), whereas Fungiplex had the lowest positivity rate (71%). No false positives were observed in negative control swabs for any assay.Conclusions: Real-time PCR is a crucial tool for the rapid and sensitive detection of C. auris , especially in clinical settings where timely identification is essential for effective patient management and infection control. Numerous PCR assays are available for this purpose; however, our study demonstrates that the sensitivity of these assays can vary significantly. The observed differences underscore the importance of establishing international reference standards and proficiency panels to enhance the accuracy and comparability of assay performance across different studies and laboratories.","PeriodicalId":18797,"journal":{"name":"Mycoses","volume":"68 5","pages":"e70065"},"PeriodicalIF":4.1000,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12048891/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Mycoses","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1111/myc.70065","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"DERMATOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

Objectives: This study aimed to systematically evaluate and compare the performance of two laboratory-developed assays (LDAs) and three commercially available real-time PCR assays for the detection of Candida auris. The analytical sensitivity, specificity and limit of detection (LOD) of each assay were assessed, alongside their clinical sensitivity in identifying C. auris colonisation.

Methods: Ten C. auris strains representing five clades, as well as genetically related yeasts, common yeast species, and dermatophytes, were used to assess assay sensitivity and cross reactivity. Clinical and environmental samples were collected from patients during an outbreak and tested with three commercial PCR assays (AurisID, Fungiplex, FungiXpert) and two LDAs (CDC LDA, EMC LDA). LOD was determined using Probit analysis. Diagnostic sensitivity was evaluated by comparing the detection rate of each individual assay to the total detection rate of all assays combined.

Results: The EMC LDA exhibited the highest analytical sensitivity, with a LOD of 8 conidia/reaction, followed by CDC LDA (16 conidia/reaction), AurisID and FungiXpert (19 conidia/reaction), and Fungiplex (596 conidia/reaction). Specificity testing revealed cross-reactivity in the CDC LDA and AurisID assays with C. pseudohaemulonii at high conidia levels, while no cross-reactivity was observed in the other assays. EMC LDA showed the highest clinical sensitivity (100%), whereas Fungiplex had the lowest positivity rate (71%). No false positives were observed in negative control swabs for any assay.

Conclusions: Real-time PCR is a crucial tool for the rapid and sensitive detection of C. auris , especially in clinical settings where timely identification is essential for effective patient management and infection control. Numerous PCR assays are available for this purpose; however, our study demonstrates that the sensitivity of these assays can vary significantly. The observed differences underscore the importance of establishing international reference standards and proficiency panels to enhance the accuracy and comparability of assay performance across different studies and laboratories.

查看原文本刊更多论文

五种实时荧光定量PCR检测耳念珠菌DNA的性能评价。

目的：本研究旨在系统地评价和比较两种实验室开发的检测方法（lda）和三种市售实时PCR检测方法检测念珠菌的性能。评估了每种检测方法的分析灵敏度、特异性和检出限（LOD），以及它们在鉴定耳念珠菌定殖方面的临床敏感性。方法：采用代表5个分支的10株金黄色葡萄球菌，以及遗传相关酵母菌、常见酵母菌和皮肤真菌，评估检测灵敏度和交叉反应性。在疫情期间从患者身上收集临床和环境样本，并使用三种商用PCR检测方法（AurisID、Fungiplex、FungiXpert）和两种LDA （CDC LDA、EMC LDA）进行检测。LOD采用Probit分析法测定。通过比较每个单独检测的检出率与所有检测的总检出率来评估诊断敏感性。结果：EMC LDA的分析灵敏度最高，LOD为8个分生菌/反应，其次为CDC LDA（16个分生菌/反应）、AurisID和FungiXpert（19个分生菌/反应）和Fungiplex（596个分生菌/反应）。特异性测试显示CDC LDA和AurisID检测与高分生孢子水平的假马龙假梭菌具有交叉反应性，而其他检测未观察到交叉反应性。EMC LDA临床敏感性最高（100%），而Fungiplex阳性率最低（71%）。阴性对照拭子在任何检测中均未观察到假阳性。结论：实时荧光定量PCR是快速、灵敏检测金黄色葡萄球菌的重要工具，特别是在临床环境中，及时识别对有效的患者管理和感染控制至关重要。许多PCR检测可用于此目的；然而，我们的研究表明，这些检测方法的敏感性可能会有很大差异。观察到的差异强调了建立国际参考标准和熟练度小组以提高不同研究和实验室测定性能的准确性和可比性的重要性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Mycoses 医学-皮肤病学

CiteScore

10.00

自引率

8.20%

发文量

143

审稿时长

6-12 weeks

期刊介绍： The journal Mycoses provides an international forum for original papers in English on the pathogenesis, diagnosis, therapy, prophylaxis, and epidemiology of fungal infectious diseases in humans as well as on the biology of pathogenic fungi. Medical mycology as part of medical microbiology is advancing rapidly. Effective therapeutic strategies are already available in chemotherapy and are being further developed. Their application requires reliable laboratory diagnostic techniques, which, in turn, result from mycological basic research. Opportunistic mycoses vary greatly in their clinical and pathological symptoms, because the underlying disease of a patient at risk decisively determines their symptomatology and progress. The journal Mycoses is therefore of interest to scientists in fundamental mycological research, mycological laboratory diagnosticians and clinicians interested in fungal infections.