Semiquantitative detection by real-time PCR of Aspergillus fumigatus in bronchoalveolar lavage fluids and tissue biopsy specimens from patients with invasive aspergillosis
Semiquantitative detection by real-time PCR of Aspergillus fumigatus in bronchoalveolar lavage fluids and tissue biopsy specimens from patients with invasive aspergillosis
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DOI：
10.1128/jcm.41.9.4304-4311.2003
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发表时间：
2003-09-01
影响因子：
9.4
通讯作者：
Salonen, J
中科院分区：
文献类型：
--
作者：
Rantakokko-Jalava, K;Laaksonen, S;Salonen, J
开发了一种实时 PCR 方法，用于检测支气管肺泡灌洗 (BAL) 液和组织活检标本中的烟曲霉线粒体 DNA (mtDNA)。该测定的分析灵敏度为每个反应一个烟曲霉分生孢子，并且该测定的线性至少比检测限高出4个数量级以上。对 66 名有侵袭性肺曲霉病 (IPA) 风险的免疫功能低下患者和 33 名免疫功能正常对照者的 BAL 液体以及 10 名免疫功能低下患者的组织活检标本进行了分析。结果与根据最近发布的共识标准建立的临床诊断相关。在来自高危患者的 81 份 BAL 液体样本中，有 16 份 (20%) 发现烟曲霉 mtDNA 阳性，在免疫功能正常对照的 33 份样本中，发现有 16 份 (3%) 呈烟曲霉 mtDNA 阳性。七名患者中的六名、四名中的两名和五名中的四名患者的 PCR 呈阳性，分别是已证实的、可能的和可能的 IPA，以及四名有风险但没有任何其他 IPA 证据的患者。通过定性检测，PCR的诊断灵敏度为73%，特异性为93%，阳性（PPV）和阴性（NPV）结果的预测值分别为73%和95%。使用阈值循环
A real-time PCR method was developed and used to detect Aspergillus fumigatus mitochondrial DNA (mtDNA) in bronchoalveolar lavage (BAL) fluids and tissue biopsy specimens. The analytical sensitivity of the assay was one A. fumigatus conidium per reaction, and the assay was linear at least over 4 orders of magnitude above the detection limit. BAL fluids from 66 immunocompromised patients at risk of invasive pulmonary aspergillosis (IPA) and 33 immunocompetent controls and tissue biopsy specimens from 10 immunocompromised patients were analyzed. The results were related to the clinical diagnosis established according to recently published consensus criteria. A. fumigatus mtDNA positivity was encountered in 16 of 81 (20%) BAL fluid specimens from patients at risk and I of 33 (3%) specimens from immunocompetent controls. PCRs were positive in six of seven, two of four, and four of five of the patients with proven, probable, and possible IPA, respectively, as well as in four patients at risk but without any other evidence of IPA. With qualitative detection, the diagnostic sensitivity of PCR was 73%, specificity was 93%, and predictive values of positive (PPV) and negative (NPV) results were 73 and 95%, respectively. Using a threshold cycle of