Real-time automated PCR for early diagnosis and monitoring of cytomegalovirus infection after bone marrow transplantation
Real-time automated PCR for early diagnosis and monitoring of cytomegalovirus infection after bone marrow transplantation
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DOI：
10.1128/jcm.38.7.2536-2542.2000
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发表时间：
2000-07-01
影响因子：
9.4
通讯作者：
Hirai, H
中科院分区：
文献类型：
--
作者：
Machida, U;Kami, M;Hirai, H
本研究的目的是评估实时自动 PCR 作为一种定量、高重复性且灵敏的方法来检测血液样本中巨细胞病毒 (CMV) DNA 的有用性。检测内和检测间精确度分别为 0.89%（小拷贝数 [L]）、1.43%（中等拷贝数 [M]）和 1.12% 大腿拷贝数 [H]）。分别为 4.46% (L)、1.51% (M) 和 2.28% (ZI)。该测定的线性度在 10 和 10(7) 拷贝/孔之间获得，最小检测限为 20 拷贝/孔。 70名健康受试者中有55人的样本被发现CMV抗体呈阳性，但其中没有检测到CMV DNA。在每个样本的定性评估中，CMV抗原血症测定结果与实时PCR测定结果一致的比例为80%（血浆样本）、79%（所有有核细胞）和86%（血液）。对于 8 名被诊断为 CMV 感染或疾病的患者，抗原血症检测中没有样本早于实时 PCR 检测呈阳性。此外，该测定的结果可以在8小时内获得，我们得出结论，实时PCR测定对于CMV感染的快速诊断和临床过程的监测是有用的。
The purpose of this study was to assess the usefulness of real-time automated PCR as a quantitative, highly reproducible, and sensitive method to detect cytomegalovirus (CMV) DNA in blood specimens. Intra- and interassay precision rates were 0.89% (small number of copies [L]), 1.43% (middle number of copies [M]), and 1.12% thigh number of copies [H]). and 4.46% (L), 1.51% (M), and 2.28% (ZI), respectively, The linearity of this assay was obtained between 10 and 10(7) copies/well, with a minimum detection limit of 20 copies/well. Specimens from 55 of 70 healthy subjects were found to be positive for CMV antibody, but CMV DNA was not detected in any of them. In the qualitative assessment of each specimen, the results of the CMV antigenemia assay and those of the real-time PCR assay agreed in 80% (plasma specimens), 79% (all nucleated cells), and 86% (blood) of the cases examined. For eight patients diagnosed as having CMV infection or disease, no sample was positive in the antigenemia assay earlier than in the real-time PCR assay. Furthermore, the results of this assay could be obtained within 8 h, We concluded that the real-time PCR assay is useful for rapid diagnosis of CMV infection and monitoring of clinical courses.