Detection of human herpesvirus 7 DNA by loop-mediated isothermal amplification
Detection of human herpesvirus 7 DNA by loop-mediated isothermal amplification
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DOI：
10.1128/jcm.42.3.1348-1352.2004
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发表时间：
2004-03-01
影响因子：
9.4
通讯作者：
Asano, Y
中科院分区：
文献类型：
--
作者：
Yoshikawa, T;Ihira, M;Asano, Y
本研究评估了环介导等温扩增 (LAMP) 的可靠性，该技术最初是为检测人类疱疹病毒 7 (HHV-7) 而开发的。尽管在 HHV-7 DNA 中检测到了 LAMP 产物，但 HHV-6 和人巨细胞病毒 DNA 均未产生产物。当使用琼脂糖凝胶电泳检测LAMP产物时，30分钟的HHV-7 LAMP反应的灵敏度达到250拷贝/管。使用浊度检测 LAMP 产物，反应 30 分钟和 60 分钟的灵敏度分别为 500 和 250 拷贝/管。在这些初步验证研究之后，通过 HHV-7 LAMP 检查了从两名原发性 HRV-7 感染患者收集的临床样本。通过使用琼脂糖凝胶电泳，可以在急性期血浆样本中检测到 HHV-7 LAMP 产物，但在两位患者的恢复期血浆样本中均未检测到 LAMP 产物。由于浊度测定的灵敏度不如琼脂糖凝胶电泳，因此在 30 分钟的 LAMP 反应后，血浆样品中无法检测到 HHV-7 LAMP 产物。经过 60 分钟的 LAMP 反应，可以在急性期血浆样本中检测到 HHV-7 LAMP 产物。
The reliability of loop-mediated isothermal amplification (LAMP), initially developed for the detection of human herpesvirus 7 (HHV-7), was evaluated in this study. Although a LAMP product was detected in HHV-7 DNA, neither HHV-6 nor human cytomegalovirus DNA produced a product. When agarose gel electrophoresis was used for the detection of LAMP products, the sensitivity of a 30-min HHV-7 LAMP reaction reached 250 copies/tube. The use of turbidity for the detection of the LAMP products gave a sensitivity of 500 and 250 copies/tube for 30- and 60-min reactions, respectively. Following these initial validation studies, clinical samples collected from two patients with primary HRV-7 infections were examined by HHV-7 LAMP. By use of agarose gel electrophoresis, HHV-7 LAMP products could be detected in acute-phase plasma samples but no LAMP product was detectable in convalescent-phase plasma samples from either patient. Since a turbidity assay is less sensitive than agarose gel electrophoresis, no HHV-7 LAMP product could be detected in plasma samples after a 30-min LAMP reaction. After a 60-min LAMP reaction, HHV-7 LAMP product could be detected in acute-phase plasma samples.