AEGiS-11IAC: Validation of a quantitative DNA PCR assay for HIV-1 in human peripheral blood mononuclear cells.

11th International AIDS Conference

Vancouver, British Columbia — July 7-12, 1996

Validation of a quantitative DNA PCR assay for HIV-1 in human peripheral blood mononuclear cells.

Int Conf AIDS 1996 Jul 7-12; 11:16 (abstract no. We.A.520)
Wathen LK, Nuorala KW, Patel RK, DeYoung DP, Re' KA, Cromie MA, Crampton DJ, Krieger KS, Greenwald CA, Freimuth WW; Pharmacia & Upjohn, Inc., Kalamazoo, MI, USA. Fax: 616-385-7219.

OBJECTIVE: To analytically and clinically validate a precise quantitative DNA PCR assay to evaluate the therapeutic responsiveness of the HIV-1 viral load in the lymphocyte compartment of infected patients.

METHODS: A quantitative HIV-1 cellular DNA assay was rigorously standardized. Standard material, control lymphocytes, specimen handling and processing, storage and assay quality control were optimized. A systematic analysis of the precision of the assay, patient sample variability, a comparison of quantitative DNA HIV-1 levels to other surrogate and viral load markers in over 1100 patients was performed. Results:This assay was linear over a greater than 2 log range and, after analyzing more than 7000 patient samples, had an intra- and inter- assay precision of 7% and 19% respectively. When HIV-1 infected patients were off antiviral therapy for at least 14 days, 99% had measurable DNA PCR levels and 85% had a viral load greater than 1000 copies/3M lymphocytes. In contrast, only 58% of these subjects had a viral burden detectable by lymphocyte culture techniques. Unlike the plasma reservoir, the median HIV-1 lymphocyte viral burden (2500 copies/3M cells) was relatively constant in patients with 300-800 CD4+ cell count. The median DNA viral load increased to approximately 6200 copies/3M cells in patients with 50-299 CD4+ cell count. Patients treated with Delavirdine (200, 300, 400 mg TID) in combination with ZDV or ddI demonstrated a dose response relationship in diminution of viral load in lymphocyte DNA.

CONCLUSIONS: The high level of precision and reproducibility of this quantitative DNA PCR assay provide an enhanced means of evaluating therapeutic drug regimens for HIV-1. These studies illustrate the distinctiveness of the HIV-1 lymphocytic viral pool and reveal the accessibility of combination therapy to this compartment.

Keywords: AEGIS, HIV-1, Viral Load, Polymerase Chain Reaction, CD4 Lymphocyte Count, DNA, DNA, Viral, DNA Primers, Human, ICA11

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