Tetali S K, Oyaizu N, Paul M, Pahwa S
Department of Pediatrics, North Shore University Hospital-Cornell University Medical College, Manhasset, NY 11030.
AIDS Res Hum Retroviruses. 1993 Jan;9(1):77-82. doi: 10.1089/aid.1993.9.77.
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.
本研究的目的是调查外周血单核细胞（PBMC）中刺激诱导的1型人类免疫缺陷病毒（HIV-1）表达上调是否能提高聚合酶链反应（PCR）的诊断敏感性。来自11名感染HIV-1的无症状成年人的PBMC在裂解细胞进行PCR之前，用植物血凝素（PHA）加佛波酯（PMA）刺激培养36小时。在所有11名研究患者中，对刺激细胞进行的PCR辅助HIV RNA扩增（RNA-PCR）强度显著高于未刺激细胞（p<0.001）。对7名患者进行了传统PCR辅助DNA扩增（DNA-PCR）与RNA-PCR的比较。细胞预先刺激后，DNA-PCR的敏感性也有所提高，尽管程度不如RNA-PCR。我们的研究结果表明，PHA和PMA预先激活细胞可显著提高PCR的敏感性。
