Quantification of human cytomegalovirus using bronchoalveolar lavage cells in pulmonary complications associated with hematologic neoplasia

Human cytomegalovirus (CMV) has been recognized as a frequent pathogen involved in interstitial pneumonia (IP), and CMV-IP is a severe and life-threatening complication in the immunocompromised patients. The use of real-time PCR in molecular diagnostics has increased to the point where it is now accepted as the gold standard for detecting a wide variety of templates including viruses. Therefore, we developed a rapid quantification system of CMV using a LightCycler in order to clarify the possible role of CMV reactivation in patients with hematologic neoplasia showing pulmonary-complications. Sixty-nine bronchoalveolar lavage fluid (BALF) specimens were obtained from consecutively treated patients showing interstitial shadow including 20 patients with hematologic neoplasia. First, we determined the viral burden in BAL cells from healthy volunteers, idiopathic interstitial pneumonia (IIP) and sarcoidosis. CMV copy numbers in samples obtained from healthy volunteers, IIP and sarcoidosis, were less than 10(2) copies per 1 mug of DNA, whether or not BAL cells were composed of high percentage of lymphocytes. Among 20 patients with hematologic neoplasia analyzed, two specimens obtained from leukemia patients with pulmonary alveolar proteinosis, two from GvHD, one with CMV interstitial pneumonia and one with Hodgkin's disease had high level of CMV viral DNA. Our results suggest that measurement of CMV genomes in BAL cells using real-time PCR may be useful not only to understand the involvement of CMV in systematic respiratory tract disease but also in management of the care of respiratory complications in hematologic neoplasia.