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A simple and sensitive method for detecting adenovirus in serum and urine
Y C Henderson1, T J Liu, G L Clayman
1Department of Head and Neck Surgery, The University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
Journal of Virological Methods
|June 17, 1998
Summary
A new method accurately detects shed adenovirus (Ad5CMV-p53) in patient samples. This sensitive technique uses polyethylene glycol precipitation and PCR, achieving high detection limits in under two days.
Area of Science:
- Virology
- Molecular Biology
- Biotechnology
Background:
- Gene therapy using viral vectors like adenovirus (Ad5CMV-p53) requires monitoring viral shedding in patients.
- Detecting low levels of shed virus in biological fluids is crucial for assessing treatment safety and efficacy.
Purpose of the Study:
- To develop a sensitive and reproducible method for detecting shed Ad5CMV-p53 in patient serum and urine.
- To establish a rapid assay with a low limit of detection for adenoviral particles.
Main Methods:
- Developed a novel protocol involving polyethylene glycol precipitation of adenoviral particles from serum or urine.
- Utilized proteinase K digestion for adenoviral DNA extraction.
- Employed polymerase chain reaction (PCR) amplification combined with Southern blot transfer for enhanced detection.
Main Results:
- Achieved a detection limit of five viral particles per 0.25 ml of serum.
- Demonstrated a detection limit of one viral particle per 0.25 ml of urine.
- The entire procedure is sensitive, reproducible, and can be completed in less than 48 hours.
Conclusions:
- The developed method offers a highly sensitive and efficient way to monitor adenoviral shedding post-gene transfer.
- This assay provides a valuable tool for clinical applications requiring the detection of low viral loads.
- The rapid turnaround time facilitates timely clinical decision-making.