TY - JOUR
T1 - Detection of Human Herpesvirus 7 DNA by Loop-Mediated Isothermal Amplification
AU - Yoshikawa, Tetsushi
AU - Ihira, Masaru
AU - Akimoto, Shiho
AU - Usui, Chie
AU - Miyake, Fumi
AU - Suga, Sadao
AU - Enomoto, Yoshihiko
AU - Suzuki, Ryota
AU - Nishiyama, Yukihiro
AU - Asano, Yoshizo
PY - 2004/3
Y1 - 2004/3
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=12144289139&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=12144289139&partnerID=8YFLogxK
U2 - 10.1128/JCM.42.3.1348-1352.2004
DO - 10.1128/JCM.42.3.1348-1352.2004
M3 - Article
C2 - 15004116
AN - SCOPUS:12144289139
SN - 0095-1137
VL - 42
SP - 1348
EP - 1352
JO - Journal of clinical microbiology
JF - Journal of clinical microbiology
IS - 3
ER -