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

Toshiya Aono, Kazuhiro Kondo, Hiroko Miyoshi, Keiko Tanaka‐Taya, Motohiro Kondo, Yuko Osugi, Junichi Hara, Shintaro Okada, Koichi Yamanishi
1998 Journal of Infectious Diseases  
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 chain reaction (PCR) methods. For NASBA detection, the b 2.7 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
more » ... d 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. Human cytomegalovirus (HCMV) is a significant cause of morbidity and mortality in immunocompromised patients, such as organ or bone marrow transplant (BMT) recipients, AIDS patients, and developing neonates [1-3]. Some antiviral drugs, such as ganciclovir [4] and foscarnet [5] , are useful for treating HCMV disease, but these drugs must be used carefully because of their side effects. Therefore, it is important to establish a sensitive system by which to monitor HCMV infection. Classical viral culture from peripheral blood mononuclear cells is not a very sensitive technique, and it is time-consuming. Serologic methods are generally useful but often inaccurate in the case of immunocompromised patients because of hyperimmune globulin therapy and defective immune responses. The HCMV antigenemia assay has been shown to have good clinical specificity for symptomatic HCMV infection [6, 7] , but it is complicated and demands a large number of cells from patients, including pediatric BMT recipients and neonates. The detection of HCMV genomic DNA by polymerase chain reaction (PCR) is highly sensitive [8-10] but not always helpful in HCMV infection because it cannot distinguish latent from active infection. mRNA is a good target for active HCMV-specific detection,
doi:10.1086/314449 pmid:9780242 fatcat:up7vzr2j45f4fmcvvte26e7nau