Preliminary Report for the Application of Nucleic Acid Sequence-Based Amplification in Detection of Human Cytomegalovirus (HCMV) Late pp67 mRNA for Diagnosis of HCMV Infection
WANTANIT PAIROJ, M.Sc.*, W ASUN CHANTRA TIT A, Ph.D.*, SOMNUEK SUNGKANUPARPH, M.D.**, CHUTATIP SRICHUNRUSAMI, B.Sc.*, PISAMAI BODHIPHALA, M.Sc.*, ASDA VIBHAGOOL, M.D.**
Affiliation : * Virology and Molecular Microbiology Laboratory, Department of Pathology, ** Department of Medicine, Faculty of Medicine, Ramathibodi Hospital. Mahidol University, Bangkok 10400, Thailand.
Human cytomegalovirus (HCMV) late pp67 mRNA expression by nucleic acid sequence- based amplification (NASBA) in patients, clinically diagnosed as possible HCMV, probable HCMV disease, and no disease, was evaluated. The RNAs were isolated from II whole-blood samples of 11 patients for the specific amplification of the pp67 mRNA. NASBA results were compared to results from PCR assay and serological assay. The HCMV pp67 mRNA could be found in 3 of 11 patients, whereas, HCMV-DNA PCR was positive in 6 of II patients. PCR assay for HCMV -DNA in plasma has proved to correlate with clinical diagnosis of HCMV infection. Only 2 patient samples of NASBA positive results coincided with HCMV -DNA PCR. However, the diagnosis of clinically relevant HCMV infection by NASBA was seen. Anti-CMV IgG titers of 1: I ,600 or over 1:1,600 were found in 2 of 3 NASBA positve cases and 5 of 6 HCMV-DNA positive cases, whereas, anti-CMV IgM were all negative. These results showed the correlation of HCMV infection detected by NASBA, PCR assay and anti-CMV IgG of the titers up to 1: I ,600. Additionally, a low antibody titer of the HIV patient could be diag- nosed by NASBA or PCR. In conclusion, pp67 mRNA NASBA appears to be a promising diag- nostic tool in analysis of HCMV infection and/or disease. Its diagnostic value should be defined in the specific group for the follow-up of immunocompromised patients, such as organ transplant recipients in future prospective studies.
Keywords : pp67 mRNA, NASBA, HCMV-DNA, PCR
