Open AccessRapid Diagnosis of Herpes Simplex Virus Infection by a Loop-Mediated Isothermal Amplification Method
Author(s) -
Yoshihiko Enomoto,
Tetsushi Yoshikawa,
Masaru Ihira,
Shiho Akimoto,
Fumi Miyake,
Chie Usui,
S. Suga,
Kayoko Suzuki,
Takashi Kawana,
Yukihiro Nishiyama,
Yoshizo Asano
Publication year - 2005
Publication title -
journal of clinical microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.349
H-Index - 255
eISSN - 1070-633X
pISSN - 0095-1137
DOI - 10.1128/jcm.43.2.951-955.2005
Subject(s) - loop mediated isothermal amplification , agarose gel electrophoresis , herpes simplex virus , agarose , microbiology and biotechnology , virology , biology , gel electrophoresis , polymerase chain reaction , hsl and hsv , virus , dna , gene , biochemistry
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.