Comparison of the quantitative competitive and semiquantitative RT-PCR methods for the determination of interferon-gamma mRNA levels in AIDS-free HIV-infected individuals

• Main Authors: Barabás É., Nagy K., Várkonyi V., Horváth A.
• Format: Article
• Language: English
• Published: Associação Brasileira de Divulgação Científica 2001-01-01
• Series: Brazilian Journal of Medical and Biological Research
• Subjects: quantitative competitive RT-PCR; semiquantitative RT-PCR; IFN-gamma mRNA; HIV
• Online Access: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2001001000006

IFN-gamma mRNA expression was evaluated in nonstimulated peripheral blood mononuclear cells (PBMC) of HIV-infected and seronegative individuals using quantitative competitive and semiquantitative RT-PCR and the sensitivity of these methods was compared. A significant correlation was found between quantitative competitive and semiquantitative RT-PCR in samples of both HIV-seronegative (P = 0.004) and HIV-infected individuals (P = 0.0004). PBMC from HIV-infected individuals presented a remarkable increase of IFN-gamma mRNA expression, as determined by both types of RT-PCR methods. Semiquantitative RT-PCR even without an internal standard is also acceptable for measuring cytokine mRNA expression, but less reliable if small amounts are quantified. Moreover, we found that increased IFN-gammamRNA expression is independent of CD4+ cell count in AIDS-free HIV-infected patients.