Development of a Sensitive DNA Assay for the AIDS Virus, HTLV-III/LAV
Annual rept. 16 Mar 1988-15 Mar 1989
WASHINGTON UNIV ST LOUIS MO
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The study was undertaken in order to devise a quantitative assay of human immunodeficiency virus type 1 HIV-1 load in patients tissues. No methods are currently available which are both sensitive and specific for this purpose. Our method utilizes blood mononuclear cell or tissue DNA. HIV-1 sequences are amplified by the primer chain amplification reaction PCR technique. Reaction products are detected and quantitated by polyacrylamide or agarose gel electrophoresis, ethidium bromide stain and densitometry. This method has proven successful using a single set or primers in detection of HIV-1 DNA sequences from 10 of 12 HIV-1 infected individuals, and 0 out of 10 uninfected individuals. Methods of quantitation using standard curves with defined amounts of HIV-1 DNA sequences and internal controls have been developed. We have developed a sensitive and specific assay from HTLV-I and HTLV-II DNA sequences using the same methodology and successfully applied it to the evaluation of blood samples from asymptomatic HTLV-I infected individuals and individuals with adult T cell leukemia-lymphoma.
- Medicine and Medical Research