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Journal of Clinical Microbiology, November 1998, p. 3323-3326, Vol. 36, No. 11
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

PCR Method for Detection of Adenovirus in Urine of Healthy and Human Immunodeficiency Virus-Infected Individuals

Marcela Echavarria,1 Michael Forman,1 John Ticehurst,1,2 J. Stephen Dumler,1 and Patricia Charache1,*

Division of Medical Microbiology, Department of Pathology, The Johns Hopkins Medical Institutions, Baltimore,1 and Microbiology Branch, Division of Clinical Laboratory Devices, Office of Device Evaluation, Center for Devices and Radiological Health, U.S. Food and Drug Administration, Rockville,2 Maryland

Received 15 May 1998/Returned for modification 22 July 1998/Accepted 20 August 1998

Adenoviruses (AdV) cause diseases that range from localized, self-limited illnesses to fatal infections in immunocompromised patients. Culture is assumed to be sensitive but requires viable virus and up to 3 weeks 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 sensitivity than obtainable by culture technology. All 18 serotypes tested were detected. Quantitatively, with optimized urine processing, AdV PCR detected 0.2 PFU/ml (serotype 11) and 10 DNA copies/ml (serotype 2). Serially collected urine samples from human immunodeficiency virus (HIV)-infected patients with concurrent cytomegalovirus retinitis were divided into three groups: AdV culture-positive samples, AdV culture-negative or bacterially contaminated samples from patients with a history of AdV culture-positive urines, and AdV culture-negative samples from patients without a history of AdV culture positivity. Urine samples from healthy adults were also tested by culture and PCR to screen for asymptomatic shedding. Amplification was assessed with and without prior DNA purification. AdV was detected by PCR in 90% of culture-positive urines (100% of unclotted samples, e.g., those culture positive after storage for PCR testing), 71% of culture-negative or bacterially contaminated urines from AdV-infected patients, and 28% from AdV culture-negative patients. Healthy volunteers were culture negative for AdV, and 96% were PCR negative. The new AdV PCR method is rapid and sensitive and can detect viral DNA in samples for which culturing is problematic. The role of AdV replication during HIV infection merits further investigation with sensitive tools such as PCR.

* Corresponding author. Mailing address: Division of Medical Microbiology, Department of Pathology, The Johns Hopkins Hospital, 600 North Wolfe St., Baltimore, MD 21287. Phone: (410) 955-5775. Fax: (410) 614-7475. E-mail: pcharach{at}pathlan.path.jhu.edu.

Journal of Clinical Microbiology, November 1998, p. 3323-3326, Vol. 36, No. 11
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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