使用实时聚合酶链反应检测血液样本以诊断侵袭性曲霉病。
Use of real-time PCR on blood samples for diagnosis of invasive aspergillosis.
作者信息
Kami M, Fukui T, Ogawa S, Kazuyama Y, Machida U, Tanaka Y, Kanda Y, Kashima T, Yamazaki Y, Hamaki T, Mori S, Akiyama H, Mutou Y, Sakamaki H, Osumi K, Kimura S, Hirai H
机构信息
Department of Hematology and Oncology, Graduate School of Medicine, University of Tokyo, Japan.
出版信息
Clin Infect Dis. 2001 Nov 1;33(9):1504-12. doi: 10.1086/323337. Epub 2001 Oct 4.
We developed a new quantitative system for diagnosis of invasive pulmonary aspergillosis (IPA) using real-time 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 -0.3+/-6.6 days, those of BDG measurement by 6.5+/-4.9 days, and those of ELISA by 2.8+/-4.1 days. Real-time PCR was sensitive for IPA diagnosis, and quantitation was accurate.
我们开发了一种使用实时自动聚合酶链反应（PCR）诊断侵袭性肺曲霉病（IPA）的新定量系统。体外检测的批内和批间精密度率分别为2.53%和2.20%，当每孔有>20个拷贝时可获得该检测方法的线性。我们检测了从122例血液系统恶性肿瘤患者中采集的323份样本，其中包括33例IPA患者和89例对照患者。血样采用酶联免疫吸附测定（ELISA）和血浆（1→3）-β-D-葡聚糖（BDG）浓度测定的PCR抗原检测方法。PCR、ELISA和BDG测量诊断IPA的敏感性分别为79%、58%和67%；特异性分别为92%、97%和84%。PCR阳性结果比计算机断层扫描提前-0.3±6.6天，比BDG测量提前6.5±4.9天，比ELISA提前2.8±4.1天。实时PCR对IPA诊断敏感，定量准确。
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