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:

• Meeting Abstracts

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:

• 00142789

UI: 102177318

From Meeting Abstracts