QUANTITATIVE DETECTION OF HUMAN CYTOMEGALOVIRUS DNA IN LUNG-TRANSPLANT RECIPIENTS

Human cytomegalovirus (HCMV) disease remains a major cause of morbidit y and mortality after lung transplantation. Currently, routine diagnos tic tests for HCMV are inefficient and insensitive or nonspecific for HCMV disease. We describe an efficient, highly sensitive, quantitative polymerase chain reaction (PCR) assay for HCMV using competitive PCR and fluorescently labeled primers, and we have used this to measure HC MV DNA load in donor and recipient tissues of six lung transplant reci pients at the time of transplantation, and 2 wk after transplantation when clinically stable. Total DNA yield was adequate for analysis in t ransbronchial biopsy, bronchoalveolar ravage, and peripheral blood leu kocytes, but the endobronchial biopsy specimens did not consistently p roduce sufficient DNA for analysis. There was a large intersubject and intrasubject variability between tissues in HCMV DNA load, with a ten dency for greater levels in lung tissue compared with BAL or periphera l blood cells. All six HCMV IgG seronegative donors or recipients were found to have HCMV DNA present. One of the three seronegative matched transplant recipients developed histopathologically proven HCMV disea se, and HCMV DNA levels were shown to increase at that time point and subsequently decrease with ganciclovir treatment. This assay will allo w prospective studies to confirm the predictive value of HCMV DNA load in donor and recipient tissues for HCMV disease.