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首页> 外文期刊>Journal of Clinical Microbiology >Detection of Human Herpesvirus 7 DNA by Loop-Mediated Isothermal Amplification
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Detection of Human Herpesvirus 7 DNA by Loop-Mediated Isothermal Amplification

机译:环介导的等温扩增检测人疱疹病毒7 DNA

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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 HHV-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.
机译:在这项研究中评估了最初为检测人疱疹病毒7(HHV-7)而开发的环介导的等温扩增(LAMP)的可靠性。尽管在HHV-7 DNA中检测到LAMP产物,但HHV-6和人巨细胞病毒DNA均未产生产物。当琼脂糖凝胶电泳用于检测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产物分别对30分钟和60分钟反应的灵敏度为500和250拷贝/管。在进行这些初步验证研究之后,通过HHV-7 LAMP检查了从两名原发性HHV-7感染患者中收集的临床样本。通过琼脂糖凝胶电泳,可以在急性期血浆样品中检测到HHV-7 LAMP产物,但在任一患者的恢复期血浆样品中均未检测到LAMP产物。由于浊度测定法不如琼脂糖凝胶电泳灵敏,因此在30分钟的LAMP反应后,血浆样品中未检测到HHV-7 LAMP产物。 LAMP反应60分钟后,可以在急性期血浆样品中检测到HHV-7 LAMP产物。

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