Rapid detection of human herpesvirus 8 DNA using loop-mediated isothermal amplification.
The reliability of a loop-mediated isothermal amplification (LAMP) method for the detection of human herpesvirus 8 (HHV-8) DNA was evaluated. Although LAMP products were produced with the DNA sample extracted from BCP-1 cells, LAMP products were not produced with the DNAs from seven other human herpesviruses. The detection limit of the HHV-8 LAMP method was 100 copies of target sequence/tube. To determine whether the HHV-8 LAMP method could be used to quantify viral DNA, threshold times, which are defined as the time (in s) it takes to reach the threshold turbidity level (0.1), were measured for the amplification of serial dilutions of a DNA plasmid containing the target sequence. The standard curve possessed a correlation coefficient of 0.9428 with a slope of -84.079 and y-intercept value of 1936.2. Additionally, an attempt was made to detect viral DNA in 17 specimens collected from Kaposi's sarcomas and two cell lines obtained from primary effusion lymphomas. HHV-8 DNA was detected in 14 of the 17 Kaposi's sarcoma tissue samples and both of the primary effusion lymphoma cell lines. Viral DNA was not detected in HHV-8 LAMP-negative samples using the real-time PCR method.
Kuhara
Tomoe
T
Department of Dermatology, Aichi Medical University School of Medicine, Aichi, Nagakute, Japan.
Yoshikawa
Tetsushi
T
Ihira
Masaru
M
Watanabe
Daisuke
D
Tamada
Yasuhiko
Y
Katano
Harutaka
H
Asano
Yoshizo
Y
Matsumoto
Yoshinari
Y
D004279:DNA, Viral / Q000032:analysis*; D019288:Herpesvirus 8, Human / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012514:Sarcoma, Kaposi / Q000175:diagnosis* / Q000821:virology; D012680:Sensitivity and Specificity
D004279:DNA, Viral