The Effect of Fresh Lymphocytes on Increased Sensitivity of HIV‐1 IsolationA Multicenter Study
作者:
Homayoon,
Farzadegan David,
Imagawa Phalguni,
Gupta Moon,
Lee Lisa,
Jacobson Alfred,
Saah Kathie,
Grovit Charles,
Rinaldo B.,
期刊:
Journal of Acquired Immune Deficiency Syndromes
(OVID Available online 1990)
卷期:
Volume 3,
issue 10
页码: 981-986
ISSN:0894-9255
年代: 1990
出版商: OVID
关键词: Lymphocytes;HIV-1;Isolation procedures.
数据来源: OVID
摘要:
SummaryA multicenter study was undertaken to determine the sensitivity and reproducibility of markers for human immunodeficiency virus type 1 (HIV −1) viral growth and the effect of various preparations of lymphocytes on the sensitivity of standard and routinely used procedures for HIV-1 isolation. In phase 1, cocultivated culture supernatants obtained from 10 HIV-1 cultures were transported to three Multicenter AIDS Cohort Study (MACS) Virology Laboratories. Three commercial HIV-p24 antigen capture (AC) tests and two reverse transcriptase (RT) assays were used to ascertain the replication of HIV-1. The Du Pont and Abbott AC assays were found to be most sensitive (85–100%), and the RT assay with 24-h incubation period had comparable sensitivity (75–100%). In phase II, the sensitivity of standard cocultivation procedure for HIV-1 isolation was compared using freshly phytohemagglutinin-P (PHA-P)-stimulated, stimulated-frozen, and frozen-thawed and then stimulated normal human peripheral blood mononuclear cells (PBMCs) as cocultivating cells. Blood samples from 13 HIV-1 infected individuals with various CD4+cell counts were cocultivated in each of the three MACS laboratories using one of the aforementioned normal PBMCs. The PHA-P-stimulated fresh normal PBMC showed a maximum isolation rate of 100% (13 of 13) with an average of 8 days to positivity. This rate of isolation was significantly greater than other rates using any one of the other PBMC preparations. These findings demonstrated that the use of freshly PHA-P stimulated PBMCs maximized HIV-1 isolation from blood when a sensitive HIV-1 p24 AC assay or RT assay with overnight incubation is employed for the detection of HIV in culture supernatant.
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