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首页> 外文期刊>Journal of Clinical Microbiology >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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Semiquantitative Detection by Real-Time PCR of Aspergillus fumigatus in Bronchoalveolar Lavage Fluids and Tissue Biopsy Specimens from Patients with Invasive Aspergillosis

机译：实时PCR半定量检测侵袭性曲霉病患者支气管肺泡灌洗液中的烟曲霉和组织活检标本

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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 1 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 <35 as a limit for positive PCR, the specificity and PPV of PCR in the diagnosis of invasive aspergillosis were 100%, but its sensitivity was only 45% and NPV was 92%. PCR was positive in tissue biopsy specimens from all patients with invasive aspergillosis caused by A. fumigatus. Semiquantitative detection of A. fumigatus mtDNA in BAL fluid may be helpful in the diagnosis of IPA. PCR is well suited for the verification of the presence of A. fumigatus in tissue biopsy specimens.
机译：开发了一种实时PCR方法，用于检测支气管肺泡灌洗液（BAL）和组织活检标本中的烟曲霉线粒体DNA（mtDNA）。该测定法的分析灵敏度为 A。每次反应都产生烟熏分生孢子，并且该分析至少在检测限以上至少四个数量级上呈线性。分析了来自66例有侵袭性肺曲霉病（IPA）风险的免疫功能低下患者的BAL液和33例具有免疫功能的对照以及10例免疫功能低下患者的组织活检标本。结果与根据最近公布的共识标准建立的临床诊断有关。 A。来自高危患者的81例BAL液体样本中有16例（20％）以及有免疫能力的对照的33例样本（3％）中有16例（20％）烟气mtDNA阳性。在分别证实，可能和可能患有IPA的患者中，分别有七分之六，四分之二和五分之四的PCR阳性，以及四名有风险但无其他IPA证据的患者，PCR阳性。通过定性检测，PCR的诊断敏感性为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<35的阈值循环作为阳性PCR的极限，PCR在诊断侵袭性曲霉病中的特异性和PPV为100％，但其敏感性仅为45％，NPV为92％。所有 A引起的侵袭性曲霉病患者的组织活检标本中PCR均为阳性。烟熏。半定量检测 A。 BAL液中的烟 mtDNA可能有助于IPA的诊断。 PCR非常适合验证 A的存在。组织活检标本中的烟熏。

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