Ryder TB, Fletcher HR, Cabanas DK, McDonough SH, Billyard ER, Fultz TJ, Chandler S, Nunomura K, Harper ME, Kacian DL; International Conference on AIDS.
Int Conf AIDS. 1990 Jun 20-23; 6: 155 (abstract no. F.A.319).
Gen-Probe, Inc. San Diego, CA. USA.
OBJECTIVE: We have developed a sensitive, homogeneous hybridization protection assay (HPA) which, when used in conjunction with recently developed nucleic acid target amplification methods, affords a simple and highly effective method for identifying HIV infection. METHODS: HIV genetic information was amplified using standard PCR or a proprietary amplification method which does not require repetitive temperature cycling. The amplification reactions were analyzed by HPA which involves a 15 min solution hybridization, a 6 min chemical selection against label attached to unhybridized probe, and chemiluminescent quantitation of hybridized label in a luminometer. RESULTS: Several sets of amplification primers were identified which can direct amplification effectively enough to permit detection of less than or equal to 10 HIV genomes in the presence of 10-15 mug human DNA. A good proportional relationship was observed between the HPA signal obtained and HIV genomic concentration over about 2-3 orders of magnitude. When known seropositive and seronegative patient specimens were tested, frequency distributions of the HPA signals showed excellent discrimination between positive and negative populations. CONCLUSION: HPA is a valuable complement to target amplification methods for detecting low titer infectious agents. The solution hybridization method is easier and inherently more reproducible than blot hybridization methods, and the numerical result obtained allows more reliable quantitation than the results of more subjective analytical methods such as exposure density.
Publication Types:
Keywords:
- Acquired Immunodeficiency Syndrome
- Biological Assay
- DNA
- DNA Primers
- DNA Probes
- HIV
- HIV Infections
- HIV Seropositivity
- Human Development
- Humans
- Nucleic Acid Hybridization
- Oligonucleotide Probes
- Polymerase Chain Reaction
- analysis
- methods
Other ID:
UI: 102184224
From Meeting Abstracts