Am. J. Respir. Crit. Care Med., Volume 156, Number 4, October 1997, 1241-1246

Quantitative Detection of Human Cytomegalovirus DNA in Lung Transplant Recipients

A.THOMAS C.  KOTSIMBOS, VINCENT SINICKAS, ERIC M. GLARE, DONALD S. ESMORE, GREGORY I. SNELL, E. HAYDN WALTERS, and TREVOR J. WILLIAMS

Department of Respiratory Medicine, Respiratory Immunology Group, Department of Medicine, Heart and Lung Transplant Service, Monash University Medical School, Alfred Hospital, Prahran; Department of Microbiology, Royal Melbourne Hospital, Parkville, Victoria, Australia

Human cytomegalovirus (HCMV) disease remains a major cause of morbidity and mortality after lung transplantation. Currently, routine diagnostic 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 HCMV DNA load in donor and recipient tissues of six lung transplant recipients at the time of transplantation, and 2 wk after transplantation when clinically stable. Total DNA yield was adequate for analysis in transbronchial biopsy, bronchoalveolar lavage, and peripheral blood leukocytes, but the endobronchial biopsy specimens did not consistently produce sufficient DNA for analysis. There was a large intersubject and intrasubject variability between tissues in HCMV DNA load, with a tendency for greater levels in lung tissue compared with BAL or peripheral 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 disease, and HCMV DNA levels were shown to increase at that time point and subsequently decrease with ganciclovir treatment. This assay will allow prospective studies to confirm the predictive value of HCMV DNA load in donor and recipient tissues for HCMV disease. Kotsimbos ATC, Sinickas V, Glare EM, Esmore DS, Snell GI, Walters EH, Williams TJ. Quantitative detection of human cytomegalovirus DNA in lung transplant recipients.

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