Detection of human herpesvirus 7 DNA by loop-mediated isothermal amplification.
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.
Department of Pediatrics, Fujita Health University School of Medicine, Toyoake, Aichi, Japan. tetsushi@fujita-hu.ac.jp
Tetsushi
T
Yoshikawa
Masaru
M
Ihira
Shiho
S
Akimoto
Chie
C
Usui
Fumi
F
Miyake
Sadao
S
Suga
Yoshihiko
Y
Enomoto
Ryota
R
Suzuki
Yukihiro
Y
Nishiyama
Yoshizo
Y
Asano
D001483:Base Sequence; D017931:DNA Primers; D004279:DNA, Viral / Q000097:blood / Q000737:chemistry / Q000235:genetics*; D005784:Gene Amplification; D016199:Herpesvirus 7, Human / Q000235:genetics* / Q000302:isolation & purification*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D019349:Roseolovirus Infections / Q000097:blood / Q000821:virology
D017931:DNA Primers; D004279:DNA, Viral