Rapid and reliable diagnosis of Human Immunodeficiency Virus (HIV) Type I that causes autoimmune deficiency syndrome (AIDS) is still important today In this study, the HIV-1 Tat (trans-activator of transcription) protein-specific RNA-aptamer (antiTat) and spectroscopic ellipsometer were preferred to increase specificity and sensitivity in the diagnosis. The ellipsometry is a well-known characterization tool for the ultra-thin films, where polarization state changes show surface deposition in terms of the ellipsometric angles, psi (Psi) and delta (Delta). Here, we reported the HIV-Tat protein detection performance of antiTat aptamers both for the spectroscopic ellipsometry (SE) and for the surface plasmon resonance enhanced total internal reflection ellipsometry (SPReTIRE), first time. Detection limits for antiTat aptamers with various configurations were in the range of nM-pM protein in the buffer solution. for instance, SPRe-TIRE configuration revealed a detection limit of 1 pM (or about 1.5 pg/mL) for HIV-Tat protein in the range of 1.0-500 nM. (C) 2019 Elsevier B.V. All rights reserved.