Elimination of toxicity and enhanced cytomegalovirus detection in cell cultures inoculated with semen from patients with acquired immunodeficiency syndrome

Abstract

Although semen is a particularly rich source of cytomegalovirus (CMV) and is useful for monitoring CMV shedding, its culturing is associated with extensive monolayer toxicity, isolation failures, and lengthy detection times. Inoculation of fractionated semen with immunoperoxidase staining of monolayers eliminated virtually all toxicity, increased isolation rates and monolayer infectivity, and greatly reduced detection times. Semen specimens (n = 73) were processed conventionally (C) or separated into supernatant (SF) and cellular pellet (PF) fractions, and 35% of C and SF inocula produced extensive toxicity. In contrast, virtually no toxicity was observed in monolayers inoculated with PF. C and SF isolation rates were 41 of 73 and 38 of 73, respectively, whereas that for PF was 51 of 73. Although monolayer infectivity at initial CMV detection was often less than 10% for C and SF, it was as much as 25% for PF. Average detection times were reduced from 13 days for C and SF to 6 days with PF and were further reduced to 3 days when PF inoculation was combined with immunoperoxidase staining. Thirty percent of specimens negative by C were positive by PF.