On the basis of sequence variation in the UL55 gene that encodes glycoprotein B (gB), human cytomegalovirus (CMV) can be classified into 4 gB genotypes. The goal of the present study was to determine the distribution of CMV gB genotypes and the effect of gB type on clinical outcomes in a cohort of immunocompromised patients, including both transplant recipients and nonrecipients. The distribution of gB genotypes was as follows: gB1, 28.9% of patients; gB2, 19.6%; gB3, 23.7%; gB4, 2.0%; and mixed infection, 25.8%. In contrast to patients infected with a single gB genotype, patients infected with multiple gB genotypes developed progression to CMV disease, had an increased rate of graft rejection, had higher CMV loads, and were significantly more often infected with other herpesviruses. The presence of multiple gB genotypes, rather than the presence of a single gB genotype, could be a critical factor associated with severe clinical manifestations in immunocompromised patients.