Kuhns M, McNamara A, Bankowski M, Kessler H, Landay A, Falk L; International Conference on AIDS.
Int Conf AIDS. 1989 Jun 4-9; 5: 300 (abstract no. T.B.P.80).
Abbott Laboratories, North Chicago, IL, USA
OBJECTIVE: Polymerase chain reaction (PCR) amplified sequences are usually evaluated by spot or Southern blot hybridization or gel electrophoresis of PCR products after restriction enzyme treatment. A novel solution hybridization assay was used as a convenient, sensitive and quantitative method for detection of HIV-I DNA and RNA following amplification by PCR. METHODS: The PCR product of a 680 base pair segment of the gag region was hybridized in solution with a high specific activity, single stranded 1251-DNA probe. The reaction was then applied to a gel column to separate hybrids from free probe in a single elution step. The hybridization assay itself has a sensitivity of 6x10(20) moles and is quantitative over 3.5 orders of magnitude in target concentration. RESULTS: The quantitative solution hybridization assay of DNA PCR products routinely detected 1-5 HIV-1 infected cells in 2.5x10(5) uninfected cells. HIV DNA was detected in 28/28 anti-HIV positive patients (including asymptomatic, acute and AIDS stages) and in 0/18 negative controls. The frequency of HIV infected cells in AIDS patients ranged from fewer than 9 (4% of patients) to more than 400 (13% of patients) per 10(6) mononuclear cells. CONCLUSION: These methods may be useful in monitoring DNA and RNA levels during anti-viral therapy.
Publication Types:
Keywords:
- Acquired Immunodeficiency Syndrome
- Base Sequence
- DNA
- DNA Primers
- DNA Probes
- Genes, gag
- HIV
- HIV Core Protein p24
- HIV Infections
- HIV Seropositivity
- HIV-1
- Hematologic Tests
- Humans
- Nucleic Acid Hybridization
- Oligonucleotide Probes
- Polymerase Chain Reaction
- RNA Probes
- Sensitivity and Specificity
- genetics
- immunology
Other ID:
UI: 102177318
From Meeting Abstracts