Rasheed S, Fiscus S, Griffith B, Kappes J, Elbeik T; International Conference on AIDS.
Int Conf AIDS. 1993 Jun 6-11; 9: 548 (abstract no. PO-B41-2478).
Univ. So. Cal.
Factors essential to a simple, reproducible quantitative HIV plasma viremia assay were evaluated: 1) assay dilution schema, 2) washing procedures, and 3) anticoagulant usage. Plasma from 20 HIV+ individuals was cultured in PHA-stimulated PBMC for 14 days. The titers ranged from < 1 to 15625 TCID. Two initial dilutions were tested 1:5 and neat followed by five 5-fold dilutions. In 91% there was no significant difference in end point titer and no false negative wells in the first dilution. Samples which were added neat to the first well were tested with or without washing 1 day post infection. Washing was performed in tubes and in 24-well plates. There were no differences in endpoint titers, with agreement in 90% of the samples. Heparinized and citrated plasma was tested in these assays. Virus could be quantitated from both, however, citrated plasma yielded more positives (51% vs 28%). Thus, the simplest, sensitive quantitative plasma assay utilizes neat citrated plasma with 5-fold dilutions and a media change on day 1 with no washing.
Publication Types:
Keywords:
- Acquired Immunodeficiency Syndrome
- CD4 Lymphocyte Count
- Culture
- Evaluation Studies
- HIV
- HIV Antibodies
- HIV Antigens
- HIV Core Protein p24
- HIV Infections
- HIV Seronegativity
- HIV Seropositivity
- Laboratory Techniques and Procedures
- Plasma
- Viremia
- ethnology
- immunology
- methods
- organization & administration
Other ID:
UI: 102205502
From Meeting Abstracts