Human Immunodeficiency Virus Type 1 RNA Detection in Peripheral Blood Mononuclear Cells by Polymerase Chain Reaction: Enhanced Sensitivity after Mitogenic Stimulation

Surya Kumari Tetali, Naoki Oyaizu, Morris Paul, Savita Pahwa

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

The aim of this study was to investigate whether stimulus-induced up-regulation of human immunodeficiency virus type 1 (HIV-1) expression in peripheral blood mononuclear cells (PBMC) could enhance the diagnostic sensitivity of the polymerase chain reaction (PCR). PBMC derived from 11 HIV-1-infected asymptomatic adults were cultured with a stimulus of phytohemagglutinin (PHA) plus phorbol 12-myristate 13-acetate (PMA) for 36 h prior to lysing the cells for PCR. In all 11 patients studied, the intensity of PCR-assisted HIV RNA amplification (RNA-PCR) performed on stimulated cells was significantly (p < 0.001) higher than that obtained on unstimulated cells. A comparison of conventional PCR-assisted DNA amplification (DNA-PCR) with that of RNA-PCR was made on seven patients. The sensitivity of DNA-PCR was also increased by prior stimulation of cells, although not to the same extent as was observed for RNA-PCR. The results of our study indicate that the sensitivity of PCR can be significantly enhanced by prior activation of cells with PHA and PMA.

Original languageEnglish (US)
Pages (from-to)77-82
Number of pages6
JournalAIDS research and human retroviruses
Volume9
Issue number1
DOIs
StatePublished - Jan 1993
Externally publishedYes

ASJC Scopus subject areas

  • Immunology
  • Virology
  • Infectious Diseases

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