Use of real-time PCR on blood samples for diagnosis of invasive aspergillosis
Use of real-time PCR on blood samples for diagnosis of invasive aspergillosis
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DOI：
10.1086/323337
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发表时间：
2001-11-01
影响因子：
11.8
通讯作者：
Hirai, H
中科院分区：
文献类型：
--
作者：
Kami, M;Fukui, T;Hirai, H
我们建立了一种新的定量诊断侵袭性肺曲霉病（IPA）的实时自动聚合酶链反应（PCR）。体外检测的批内和批间精密度分别为2.53%和2.20%，当拷贝数> 20个/孔时，该方法具有良好的线性。我们检查了122例血液系统恶性肿瘤患者的323份样本，包括33例IPA患者和89例对照患者。采用酶联免疫吸附试验（ELISA）对血样进行PCR抗原检测，并测定血浆（1->3）-β-D-葡聚糖（BDG）浓度。PCR、ELISA和BDG检测诊断IPA的敏感性分别为79%、58%和67%，特异性分别为92%、97%和84%。PCR的阳性结果比计算机断层扫描的阳性结果早几天，比BDG测量的阳性结果早几天，比ELISA的阳性结果早2.8 +/- 4.1天。荧光定量PCR诊断IPA灵敏度高，定量准确。
We developed a new quantitative system for diagnosis of invasive pulmonary aspergillosis (IPA) using realtime automated polymerase chain reaction (PCR). Intra-assay and interassay precision rates for in vitro examination were 2.53% and 2.20%, respectively, and the linearity of this assay was obtained when there were > 20 copies/well. We examined 323 samples taken from 122 patients with hematological malignancies, including 33 patients with IPA and 89 control patients. Blood samples were subjected to PCR antigen detection methods, using enzyme-linked immunosorbent assay (ELISA) and determination of plasma (1-->3)-beta -D-glucan (BDG) concentration. The sensitivities of PCR, ELISA, and BDG measurement for diagnosis of IPA were 79%, 58%, and 67%, respectively; the specificities were 92%, 97%, and 84%. Positive findings on PCR preceded those of computed tomography by days, those of BDG measurement by days, and those of ELISA -0.3 +/- 6.6 by 2.8 +/- 4.1 days. Real-time PCR was sensitive for IPA diagnosis, and quantitation was accurate.