Abstract. É. Barabás, K. Nagy, V. Várkonyi and A. Horváth. National Institute of Dermato-Venereology, Budapest, Hungary

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1 Brazilian Journal of Medical and Biological Research () 34: 7-75 Comparison of the quantitative competitive and semiquantitative RT-PCR methods ISSN -879X Short Communication 7 Comparison of the quantitative competitive and semiquantitative RT-PCR methods for the determination of interferon-gamma mrna levels in AIDS-free HIV-infected individuals É. Barabás, K. Nagy, V. Várkonyi and A. Horváth National Institute of Dermato-Venereology, Budapest, Hungary Correspondence É. Barabás National Institute of Dermato-Venereology Mária utca. 4 H-85 Budapest Hungary Fax: Bareva@bor.sote.hu Received November 7, Accepted July 5, Abstract g g g Key words Quantitative competitive RT-PCR Semiquantitative RT-PCR IFN-g mrna HIV g g g Braz J Med Biol Res 34()

2 7 É. Barabás et al. g g g g g g g g g g g g g Braz J Med Biol Res 34()

3 Comparison of the quantitative competitive and semiquantitative RT-PCR methods 73 g g g g g g g.e+ A P<. P<. P<. P =. P =. (NS) P =.3 B.E-4.E C HC HIV >5 HIV = 3 HIV < HC HIV >5 HIV = 3 HIV < r =.79* (95% confidence interval) P =.4* r =.83** P =.** D r =.65* (95% confidence interval) P =.4* r =.66** P =.3** -.5 e-7 e-6 e-5 IFN-g :ß-actin (M/M) -.5 e-5 e-4... IFN-g :ß-actin (M/M) Figure. Elevated interferon-g mrna levels in AIDS-free HIV-infected individuals. A, IFN-g :ß-actin mrna ratios (ratio of molar concentrations = M/M) in HIV-seronegative (healthy controls, HC) and HIV-infected patients by quantitative competitive RT-PCR. B, IFN-g :ß-actin mrna ratios (ratio in densitometric units = RDU) for HC and HIV-infected patients evaluated by semiquantitative RT-PCR. HIV-seropositive patients are divided into three groups based on CD4+ cell counts. The Mann-Whitney U-test was used for group comparisons. Significant differences between HIV-infected and HC groups are indicated. Correlation between quantitative competitive RT-PCR and semiquantitative RT-PCR IFN-g :ß-actin mrna ratios for HC samples (C) and HIV-infected samples (D). Pearson product-moment (*) and Spearman rank (**) correlations were used for the calculations (C,D). Each individual is indicated as a single point (A-D). Braz J Med Biol Res 34()

4 74 É. Barabás et al. A B C E+.E-4.E-8 IFN-g :ß-actin (RDU) E+.E-4.E-8 Sept. 996 May 999.E+.E-4.E-8 D E F.E+.E+.E+ 995 March E-4.E Nov. 997.E-4.E-8 IFN-g :ß-actin (RDU) Sept March.E-4.E-8 Figure. Longitudinal study of the IFN-g :ß-actin mrna ratio evaluated by quantitative competitive RT-PCR (ratio of molar concentrations = M/M) (lozenges) and semiquantitative RT-PCR (ratio of densitometric units = RDU) (squares): A-C, three HIV-seronegative individuals; D-F, three HIV-infected patients. g g g g g g g g Braz J Med Biol Res 34()

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