PCR METHOD FOR DETECTION OF ADENOVIRUS IN URINE OF HEALTHY AND HUMAN IMMUNODEFICIENCY VIRUS-INFECTED INDIVIDUALS

Adenoviruses (AdV) cause diseases that range from localized, self-limi ted illnesses to fatal infections in immunocompromised patients. Cultu re is assumed to be sensitive but requires viable virus and up to 3 we eks for detection, and it can be inhibited by bacterial contamination, A new PCR method amplifying a region of the hexon gene was developed in order to detect AdV in urine more rapidly and with greater sensitiv ity than obtainable by culture technology. All 18 serotypes tested wer e detected, Quantitatively, with optimized urine processing, AdV PCR d etected 0.2 PFU/ml (serotype 11) and 10 DNA copies/ml (serotype 2), Se rially collected urine samples from human immunodeficiency virus (HN)- infected patients with concurrent cytomegalovirus retinitis were divid ed into three groups: AdV culture-positive samples, AdV culture-negati ve or bacterially contaminated samples from patients with a history of AdV culture-positive urines, and AdV culture-negative samples from pa tients without a history of AdV culture positivity. Urine samples from healthy adults were also tested by culture and PCR to screen for asym ptomatic shedding. Amplification was assessed with and without prior D NA purification. AdV was detected by PCR in 90% of culture-positive ur ines (100% of unclotted samples, e.g,, those culture positive after st orage for PCR testing), 71% of culture-negative or bacterially contami nated urines from AdV-infected patients, and 28% from AdV culture-nega tive patients. Healthy volunteers were culture negative for AdV, and 9 6% were PCR negative, The new AdV PCR method is rapid and sensitive an d can detect viral DNA in samples for which culturing is problematic. The role of AdV replication during HIV infection merits further invest igation with sensitive tools such as PCR.