title: Rapid diagnosis of herpes simplex virus infection by a loop-mediated isothermal amplification method.
abstract: 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.
authors.affiliation: Department of Pediatrics, Fujita Health University School of Medicine, Toyoake, Japan 4701192.
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Enomoto
authors.forename: Tetsushi
authors.initials: T
authors.lastname: Yoshikawa
authors.forename: Masaru
authors.initials: M
authors.lastname: Ihira
authors.forename: Shiho
authors.initials: S
authors.lastname: Akimoto
authors.forename: Fumi
authors.initials: F
authors.lastname: Miyake
authors.forename: Chie
authors.initials: C
authors.lastname: Usui
authors.forename: Sadao
authors.initials: S
authors.lastname: Suga
authors.forename: Kayoko
authors.initials: K
authors.lastname: Suzuki
authors.forename: Takashi
authors.initials: T
authors.lastname: Kawana
authors.forename: Yukihiro
authors.initials: Y
authors.lastname: Nishiyama
authors.forename: Yoshizo
authors.initials: Y
authors.lastname: Asano
mesh_terms: 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
chemical_list: D004279:DNA, Viral