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

• Published in: Journal of Clinical Microbiology Vol. 43; no. 2; pp. 951 - 955
• Main Authors: Enomoto, Yoshihiko, Yoshikawa, Tetsushi, Ihira, Masaru, Akimoto, Shiho, Miyake, Fumi, Usui, Chie, Suga, Sadao, Suzuki, Kayoko, Kawana, Takashi, Nishiyama, Yukihiro, Asano, Yoshizo
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01.02.2005
• Subjects: Biological and medical sciences; DNA, Viral - analysis; Fundamental and applied biological sciences. Psychology; Herpes Genitalis - diagnosis; Herpes Genitalis - virology; Herpes Simplex - diagnosis; Herpes Simplex - virology; Herpesvirus 1, Human - genetics; Herpesvirus 1, Human - isolation & purification; Herpesvirus 2, Human - genetics; Herpesvirus 2, Human - isolation & purification; Human viral diseases; Humans; Infectious diseases; Medical sciences; Microbiology; Nucleic Acid Amplification Techniques - methods; Sensitivity and Specificity; Stomatitis, Herpetic - diagnosis; Stomatitis, Herpetic - virology; Time Factors; Viral diseases; Viral diseases with cutaneous or mucosal lesions and viral diseases of the eye; Virology; Infection; Amplification; Microbiology; Viral disease; Herpes; Diagnosis; Method
• ISSN: 0095-1137; 1098-660X
• DOI: 10.1128/JCM.43.2.951-955.2005

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.