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首页> 外文期刊>Journal of Clinical Microbiology >Characterization and Comparison of Galactomannan Enzyme Immunoassay and Quantitative Real-Time PCR Assay for Detection of Aspergillus fumigatus in Bronchoalveolar Lavage Fluid from Experimental Invasive Pulmonary Aspergillosis
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Characterization and Comparison of Galactomannan Enzyme Immunoassay and Quantitative Real-Time PCR Assay for Detection of Aspergillus fumigatus in Bronchoalveolar Lavage Fluid from Experimental Invasive Pulmonary Aspergillosis

机译：半乳甘露聚糖酶免疫测定和实时荧光定量PCR分析检测和比较实验性侵袭性肺曲霉病中支气管肺泡灌洗液中烟曲霉的特性

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Bronchoalveolar lavage (BAL) is widely used for evaluation of patients with suspected invasive pulmonary aspergillosis (IPA). However, the diagnostic yield of BAL for detection of IPA by culture and direct examination is limited. Earlier diagnosis may be facilitated by assays that can detect Aspergillus galactomannan antigen or DNA in BAL fluid. We therefore characterized and compared the diagnostic yields of a galactomannan enzyme immunoassay (GM EIA), quantitative real-time PCR (qPCR), and quantitative cultures in experiments using BAL fluid from neutropenic rabbits with experimentally induced IPA defined as microbiologically and histologically evident invasion. The qPCR assay targeted the rRNA gene complex of Aspergillus fumigatus. The GM EIA and qPCR assay were characterized by receiver operator curve analysis. With an optimal cutoff of 0.75, the GM EIA had a sensitivity and specificity of 100% in untreated controls. A decline in sensitivity (92%) was observed when antifungal therapy (AFT) was administered. The optimal cutoff for qPCR was a crossover of 36 cycles, with sensitivity and specificity of 80% and 100%, respectively. The sensitivity of qPCR also decreased with AFT to 50%. Quantitative culture of BAL had a sensitivity of 46% and a specificity of 100%. The sensitivity of quantitative culture decreased with AFT to 16%. The GM EIA and qPCR assay had greater sensitivity than culture in detection of A. fumigatus in BAL fluid in experimentally induced IPA (P ± 0.04). Use of the GM EIA and qPCR assay in conjunction with culture-based diagnostic methods applied to BAL fluid could facilitate accurate diagnosis and more-timely initiation of specific therapy.
机译：支气管肺泡灌洗（BAL）被广泛用于评估可疑的侵袭性肺曲霉病（IPA）患者。但是，通过培养和直接检查来检测IPA的BAL的诊断率有限。可以检测BAL液中的<曲>曲霉半乳甘露聚糖抗原或DNA的测定法可以促进早期诊断。因此，我们在使用中性粒细胞减少症兔的BAL液与实验诱导的IPA定义为微生物学和组织学上明显侵袭的实验中，对半乳甘露聚糖酶免疫分析（GM EIA），定量实时PCR（qPCR）和定量培养的诊断产量进行了表征和比较。 qPCR检测针对的是烟曲霉的rRNA基因复合体。 GM EIA和qPCR分析通过接收者操作者曲线分析来表征。最佳截止值为0.75，GM EIA在未经处理的对照中的敏感性和特异性为100％。进行抗真菌治疗（AFT）时，发现敏感性下降（92％）。 qPCR的最佳临界值为36个循环，灵敏度和特异性分别为80％和100％。 qPCR的灵敏度也随着AFT降低到50％。 BAL的定量培养灵敏度为46％，特异性为100％。定量培养的灵敏度随AFT降低至16％。 GM EIA和qPCR测定法在检测 A方面具有比培养更高的灵敏度。实验诱导的IPA（ P ±0.04）的BAL液中的烟气。将GM EIA和qPCR测定法与应用于BAL液的基于培养物的诊断方法结合使用，可有助于准确诊断和更及时地开始特定治疗。

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