Development of a Sensitive DNA Assay for the AIDS Virus, HTLV-III/LAV
Final rept. 16 Mar 1987-30 Sep 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 methods utilizes blood mononuclear cell or tissue DNA. HIV-1 sequences are amplified by the primer chain amplification reaction PCR technique. The 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 of primers in detection of HIV-1 DNA sequences from 10 or 12 HIV-1 infected individuals, and 0 of 10 uninfected individuals. Methods of quantitation using standard curves with defined amounts of HIV-1 DNA sequences and internal controls have been developed. Preliminary experiments are reported applying this method to quantitation of HIV sequences in more than 50 patients. In addition, we have developed a sensitive and specific assay for HTLV-I and HTLV-II DNA sequences using the same methodology and we have 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