Abstract
For rapid detection of an active stage of humancytomegalovirus (HCMV) infection, the polymerase chain reaction (PCR) technique was used to amplify CMV-DNA existing in cells of patients after bone marrow transplantation (BMT). DNAs were extracted from peripheral blood leukocytes, bone marrow cells, urine sedimentation cells, bronchoalveolar lavage fluid cells, and salivary cells using a DNA isolation kit and 0.1-1 μg of DNA was used for PCR. PCR-primers had CMV-specific sequences corresponding to the HindIII V-region of CMV, and PCR-amplified bands were detected by agarose gel electrophoresis. Therefore, in order to detect CMV-DNA, four hours were required after sample collection.
All samples found to be CMV-positive by PCR were also positive by both direct immuno-peroxidase staining and CMV isolation using cultivation. Although approximatery 90% of adults are reported to have latent CMV infection, no bands were amplified from samples obtained from normal adults or from patients before BMT. Since interstitial pneumonia caused by CMV is highly lethal for post-BMT patients, we periodically screened for active CMV infection in these patients by PCR. When patients were found to be CMV-positive, ganciclovir was adminstrated until CMVDNA was no longer detected by PCR. Twelve post-BMT patients were found to have active CMV infection without symptoms, and all of them survived after rapid diagnosis followed by ganciclovir administration. Consequently, the rapid diagnostic method described here was valuable for determining whether ganciclovir should be prescribed for these patients.
