Detection of human herpesvirus 7 DNA by loop-mediated isothermal amplification

被引:57
|
作者
Yoshikawa, T [1 ]
Ihira, M
Akimoto, S
Usui, C
Miyake, F
Suga, S
Enomoto, Y
Suzuki, R
Nishiyama, Y
Asano, Y
机构
[1] Fujita Hlth Univ, Sch Med, Dept Pediat, Toyoake, Aichi 4701192, Japan
[2] Fujita Hlth Univ Coll, Dept Med Informat Technol, Toyoake, Aichi, Japan
[3] Nagoya Univ, Grad Sch Med, Dept Virol, Nagoya, Aichi, Japan
关键词
D O I
10.1128/JCM.42.3.1348-1352.2004
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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.
引用
收藏
页码:1348 / 1352
页数:5
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