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Journal of Clinical Microbiology, February 2005, p. 951-955, Vol. 43, No. 2
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.2.951-955.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Rapid Diagnosis of Herpes Simplex Virus Infection by a Loop-Mediated Isothermal Amplification Method

Department of Pediatrics, Fujita Health University School of Medicine,¹ Department of Medical Information Technology, Fujita Health University College, Toyoake,² Department of Dermatology, Central Hospital of Tokai Medical Institute, Tokai,³ Department of Virology, Nagoya University Graduate School of Medicine, Nagoya, Aichi,⁵ Department of Obstetrics and Gynecology, University Hospital, Mizonokuchi, Teikyo University School of Medicine, Kawasaki, Kanagawa, Japan⁴

Received 1 March 2004/ Returned for modification 3 May 2004/ Accepted 12 October 2004

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.

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