Direct detection of human herpesvirus 6 DNA in serum by the loop-mediated isothermal amplification method

• Published in: Journal of clinical virology Vol. 39; no. 1; pp. 22 - 26
• Main Authors: Ihira, Masaru, Akimoto, Shiho, Miyake, Fumi, Fujita, Ayano, Sugata, Ken, Suga, Sadao, Ohashi, Masahiro, Nishimura, Naoko, Ozaki, Takao, Asano, Yoshizo, Yoshikawa, Tetsushi
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.05.2007; Elsevier Science
• Subjects: Action of physical and chemical agents; Allergy and Immunology; Biological and medical sciences; DNA Primers; DNA, Viral - blood; Exanthem subitum; Exanthema Subitum - blood; Exanthema Subitum - virology; Female; Fever - blood; Fever - virology; Fundamental and applied biological sciences. Psychology; Herpesvirus 6, Human - genetics; HHV-6; Human herpesvirus 6; Human viral diseases; Humans; Infant; Infectious Disease; Infectious diseases; LAMP; Male; Medical sciences; Microbiology; Miscellaneous; Nucleic Acid Amplification Techniques - methods; Sensitivity and Specificity; Serum; Viral diseases; Virology; human herpesvirus 6; HHV-6; Serum; LAMP; Exanthem subitum; loop-mediated isothermal amplification; PBMCs; peripheral blood mononuclear cell; PCR; polymerase chain reaction; Amplification; Skin disease; Microbiology; Exanthema; Method; Detection; Human herpesvirus 6; Virology
• ISSN: 1386-6532; 1873-5967
• DOI: 10.1016/j.jcv.2007.02.001

Abstract Background A more rapid and easier method is needed for monitoring human herpesvirus 6 (HHV-6) infections. The loop-mediated isothermal amplification method (LAMP) can detect viral DNA with high specificity, efficiency, and speed under isothermal conditions. LAMP requires only simple equipment that is available in hospital laboratories. Objectives We evaluated LAMP as a means of detecting HHV-6 DNA directly from patients’ sera. Results The sensitivity of the HHV-6 LAMP protocol without heat denaturation was 1000 copies/tube; with heat denaturation 10 copies/tube were detected. Three hundred serum samples from children with fever were analyzed. Using HHV-6 isolation as a definition of HHV-6 infection, the sensitivity, specificity, positive predictive value, and negative predictive value of the HHV-6 LAMP method without DNA extraction were 95.5%, 95.2%, 94.0%, and 96.4%, respectively. Conclusion Direct detection of HHV-6 DNA in serum with a modified HHV-6 LAMP could be used for rapid diagnosis of exanthem subitum (ES).