Toshiya Aono

Monitoring of Human Cytomegalovirus Infections in Pediatric Bone Marrow Transplant Recipients by Nucleic Acid Sequence-Based Amplification

In the diagnosis of human cytomegalovirus (HCMV) infection, it is very important to distinguish symptomatic from asymptomatic infection. The nucleic acid sequence-based amplification (NASBA) technique was compared with single and nested polymerase chainreaction (PCR) methods. For NASBA detection, the beta2.7 transcript was chosen as a target because of its abundant active HCMV-specific expression. Of 20 pediatric bone marrow transplant (BMT) recipients, 8 developed HCMV-related clinical symptoms. The clinical sensitivities and specificities were 50% and 100% for single PCR, 100% and 67% for nested PCR, and 100% and 83% for NASBA, respectively. Follow-up of HCMV infections in pediatric BMT recipients showed that NASBA could both detect viral transcript prior to the onset of clinical symptoms and reflect clinical improvement due to antiviral therapy. These data suggest that NASBA should be useful for both predicting HCMV disease development and monitoring the effect of antiviral therapy.

Detection of Human Alpha-herpesvirus DNA using Consensus Primers and Specific Probes

A simple and rapid diagnostic assay system was developed for detecting and identifying human alpha-herpesviruses, such as herpes simplex virus types 1 and 2 and varicella-zoster virus, by polymerase chain reaction. This system was based on an amplification step, using primers that bind the DNA polymerase consensus sequence of alpha-human herpesviruses, and a detection step, using non-radioactive virus-specific probes. This method could be used to amplify any human alpha-herpesviruses, and each virus-specific probe was highly specific for identification of the amplified product. This system is readily applicable for implementation in the clinical laboratory.

Herpesvirus DNA in peripheral blood mononuclear cells of some patients with Meniere's disease.

Herpes simplex virus‐1 (HSV) or varicella zoster virus (VZV) DNA was detected by nested polymerase chain reaction in peripheral blood mononuclear cells of patients with Menieres disease (one of 28 patients for HSV‐1,2 of 28 patients for VZV) during acute illness (within 5 days after onset). On the other hand, neither HSV‐1 DNA or VZV DNA was detected in PBMCs of 50 age‐ and sex‐matched healthy individuals and 50 pregnant women. These findings may imply that reactivation of HSV‐1 or VZV may be associated with the development of some cases of Menieres disease.