Loop-mediated isothermal amplification for discriminating between human herpesvirus 6 A and B.
Genotyping of human herpesvirus 6 (HHV-6) is important clinically, particularly for the diagnosis of neurological diseases. The objective of this study was to establish a rapid HHV-6 genotyping method using the loop-mediated isothermal amplification (LAMP) method. An AccI site is located in the target sequence of HHV-6 B, but not in HHV-6 A. LAMP products were digested with the AccI enzyme and then separated by agarose gel electrophoresis to differentiate the digest pattern of the two variants. The fragment patterns were clearly different between HHV-6 A and B. In order to evaluate the reliability of this HHV-6 genotyping method for use in the clinical laboratory, serum samples from 20 patients with either primary HHV-6 infection or viral reactivation were collected and analyzed. HHV-6 DNA was amplified directly from the serum samples and all 20 LAMP products were positive for HHV-6 B.
Ihira
Masaru
M
Department of Medical Information Technology, Fujita Health University College, Toyoake, Aichi, Japan.
Ohta
Akane
A
Sugata
Ken
K
Suga
Sadao
S
Asano
Yoshizo
Y
Yoshikawa
Tetsushi
T
D004279:DNA, Viral / Q000235:genetics* / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D005838:Genotype; D015654:Herpesvirus 6, Human / Q000145:classification* / Q000302:isolation & purification*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length
D004279:DNA, Viral; D015252:Deoxyribonucleases, Type II Site-Specific; C067488:GTMKAC-specific type II deoxyribonucleases