Fréalle E, Decrucq K, Botterel F, Bouchindhomme B, Camus D, Dei-Cas E, Costa J M, Yakoub-Agha I, Bretagne S, Delhaes L
EA 3609, Département de Parasitologie-Mycologie, Faculté de Médecine, Pôle de Microbiologie, CHRU de Lille, Université de Lille 2, Laboratoire d'Ecologie du Parasitisme, Institut Pasteur de Lille, Lille, France.
Eur J Clin Microbiol Infect Dis. 2009 Mar;28(3):223-32. doi: 10.1007/s10096-008-0616-1. Epub 2008 Sep 2.
In order to improve invasive pulmonary aspergillosis (IPA) diagnosis, a real-time polymerase chain reaction (PCR) assay detecting Aspergillus spp. was developed. Its detection limit reached 2-20 conidia. The retrospective evaluation on 64 bronchoalveolar lavage (BAL) fluids from 57 patients at risk for IPA, including 20 probable and five proven IPA patients, revealed a 88% or 38% sensitivity in direct examination (DE)/culture-positive or culture-negative BAL, respectively, whereas galactomannan (GM) sensitivity reached 88% or 58%, respectively. Influence on the Aspergillus-PCR yield of BAL fluid volume, cellular count and DNA content (evaluated by human beta-globin quantification) was assessed. Significantly higher beta-globin levels were detected in Aspergillus PCR-positive (median 5,112 pg/microl) than negative (median 1,332 pg/microl) BAL fluids, suggesting that the beta-globin level could reflect BAL yields and DNA extraction. Using beta-globin for the interpretation of fungal PCR could improve the negative predictive value of this test.
为了改善侵袭性肺曲霉病（IPA）的诊断，开发了一种检测曲霉属的实时聚合酶链反应（PCR）检测方法。其检测限达到2 - 20个分生孢子。对57例有IPA风险患者的64份支气管肺泡灌洗（BAL）液进行回顾性评估，其中包括20例可能的IPA患者和5例确诊的IPA患者，结果显示在直接检查（DE）/培养阳性或培养阴性的BAL中，敏感性分别为88%或38%，而半乳甘露聚糖（GM）的敏感性分别达到88%或58%。评估了BAL液体积、细胞计数和DNA含量（通过人β-珠蛋白定量评估）对曲霉PCR产量的影响。在曲霉PCR阳性的BAL液（中位数5,112 pg/μl）中检测到的β-珠蛋白水平显著高于阴性（中位数1,332 pg/μl）的BAL液，这表明β-珠蛋白水平可以反映BAL产量和DNA提取情况。使用β-珠蛋白来解释真菌PCR结果可以提高该检测的阴性预测值。
