Rapid diagnosis of herpes simplex virus infection by a loop-mediated isothermal amplification method.
Primers for herpes simplex virus type 1 (HSV 1)-specific loop-mediated isothermal amplification (LAMP) method amplified HSV-1 DNA, while HSV-2-specific primers amplified only HSV-2 DNA; no LAMP products were produced by reactions performed with other viral DNAs. The sensitivities of the HSV-1- and HSV-2-specific LAMP methods, determined by agarose gel electrophoresis, reached 500 and 1,000 copies/tube, respectively. The turbidity assay, however, determined the sensitivity of the HSV-1- and HSV-2-specific LAMP methods to be 1,000 and 10,000 copies/tube, respectively. After initial validation studies, 18 swab samples (in sterilized water) collected from patients with either gingivostomatitis or vesicular skin eruptions were examined. HSV-1 LAMP products were detected by agarose gel electrophoresis in the 10 samples that also demonstrated viral DNA detection by real-time PCR. Nine of these 10 samples exhibited HSV-1 LAMP products by turbidity assay. Furthermore, both the agarose gel electrophoresis and the turbidity assay directly detected HSV-1 LAMP products in 9 of the 10 swab samples collected in sterilized water. Next, we examined the reliability of HSV type-specific LAMP for the detection of viral DNA in clinical specimens (culture medium) collected from genital lesions. HSV-2 was isolated from all of the samples and visualized by either agarose gel electrophoresis or turbidity assay.
Enomoto
Yoshihiko
Y
Department of Pediatrics, Fujita Health University School of Medicine, Toyoake, Japan 4701192.
Yoshikawa
Tetsushi
T
Ihira
Masaru
M
Akimoto
Shiho
S
Miyake
Fumi
F
Usui
Chie
C
Suga
Sadao
S
Suzuki
Kayoko
K
Kawana
Takashi
T
Nishiyama
Yukihiro
Y
Asano
Yoshizo
Y
D004279:DNA, Viral / Q000032:analysis; D006558:Herpes Genitalis / Q000175:diagnosis* / Q000821:virology; D006561:Herpes Simplex / Q000175:diagnosis* / Q000821:virology; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000302:isolation & purification*; D018258:Herpesvirus 2, Human / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D013283:Stomatitis, Herpetic / Q000175:diagnosis / Q000821:virology; D013997:Time Factors
D004279:DNA, Viral