A simple and sensitive method for detecting adenovirus in serum and urine.
A method to detect shed virus in patients' serum and urine was developed following Ad5CMV-p53 gene transfer via direct tumor injections. The procedure differs from those reported previously in that it first uses polyethylene glycol to precipitate adenoviral particles from patient serum or urine. Adenoviral DNA is then extracted following proteinase K digestion. Finally, polymerase chain reaction (PCR) amplification followed by Southern blot transfer are employed to enhance the limit of detection to less than ten viral particles. The detection limit in a 0.25-ml sample is five viral particles in serum and one viral particle in urine. This procedure is sensitive, reproducible, and can be completed in less than 2 days.
Henderson
Y C
YC
Department of Head and Neck Surgery, The University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
Liu
T J
TJ
Clayman
G L
GL
D000260:Adenoviruses, Human / Q000235:genetics / Q000302:isolation & purification*; D001769:Blood / Q000821:virology*; D015139:Blotting, Southern; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D011092:Polyethylene Glycols; D016133:Polymerase Chain Reaction; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D014556:Urine / Q000821:virology*; D017201:Virus Shedding
D004279:DNA, Viral; D011933:Reagent Kits, Diagnostic; D011092:Polyethylene Glycols