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一种用于检测重症监护病房(ICU)有念珠菌血症风险患者血样中五种菌种的实时聚合酶链反应(PCR)检测方法的性能评估

Performance of a Real-Time PCR Assay for the Detection of Five Species in Blood Samples from ICU Patients at Risk of Candidemia.

作者信息

Felix Gabriel N, de Freitas Vera L T, da Silva Junior Afonso R, Magri Marcello M C, Rossi Flavia, Sejas Odeli N E, Abdala Edson, Malbouisson Luiz M S, Guimarães Thais, Benard Gil, Del Negro Gilda M B

机构信息

Laboratory of Medical Mycology (LIM 53), Instituto de Medicina Tropical, Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo (HC-FMUSP), São Paulo 05403-000, Brazil.

Laboratório de Investigação Médica em Imunologia (LIM 48), Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo (HC-FMUSP), São Paulo 05403-000, Brazil.

出版信息

J Fungi (Basel). 2023 May 31;9(6):635. doi: 10.3390/jof9060635.

Abstract

The gold standard for diagnosing invasive candidiasis still relies on blood cultures, which are inefficient and time-consuming to analyze. We developed an in-house qPCR assay to identify the 5 major species in 78 peripheral blood (PB) samples from ICU patients at risk of candidemia. Blood cultures and (1,3)-β-D-glucan (BDG) testing were performed concurrently to evaluate the performance of the qPCR. The qPCR was positive for DNA samples from all 20 patients with proven candidemia (positive PB cultures), showing complete concordance with species identification in blood cultures, except for detection of dual candidemia in 4 patients, which was missed by blood cultures. Additionally, the qPCR detected species in six DNA samples from patients with positive central venous catheters blood (CB) but negative PB cultures. BDG values were similarly high in these six samples and the ones with proven candidemia, strongly suggesting the diagnosis of a true candidemia episode despite the negative PB cultures. Samples from patients neither infected nor colonized yielded negative results in both the qPCR and BDG testing. Our qPCR assay was at least as sensitive as blood cultures, but with a shorter turnaround time. Furthermore, negative results from the qPCR provided strong evidence for the absence of candidemia caused by the five major species.

摘要

侵袭性念珠菌病的诊断金标准仍然依赖于血培养,而血培养分析效率低下且耗时。我们开发了一种内部定量聚合酶链反应(qPCR)检测方法,用于鉴定78例有念珠菌血症风险的重症监护病房(ICU)患者外周血(PB)样本中的5种主要念珠菌属。同时进行血培养和(1,3)-β-D-葡聚糖(BDG)检测,以评估qPCR的性能。对于所有20例确诊为念珠菌血症(PB培养阳性)的患者的DNA样本,qPCR均呈阳性,除4例患者的双念珠菌血症在血培养中漏检外,与血培养中的菌种鉴定完全一致。此外,qPCR在6例中心静脉导管血(CB)阳性但PB培养阴性的患者的DNA样本中检测到念珠菌属。这6个样本以及确诊为念珠菌血症的样本中的BDG值同样很高,强烈提示尽管PB培养阴性,但仍可诊断为真正的念珠菌血症发作。未感染也未定植的患者样本在qPCR和BDG检测中均产生阴性结果。我们的qPCR检测方法至少与血培养一样敏感,但周转时间更短。此外,qPCR的阴性结果为不存在由5种主要念珠菌属引起的念珠菌血症提供了有力证据。

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