研究目的
Investigating the colorimetric detection of HIV-1 Tat protein using split aptamer and gold nanoparticles.
研究成果
Colorimetric detection of HIV-1 Tat protein has been successfully presented by using the split aptamer for the first time. This convenient and cost-effective method has led its way for early detection of HIV by using protein as the target, which rivals the current detection method of using antibody which needed a longer time to be detected. The SPR mechanism in GNP was exploited to perform a label free, simple and time efficient detection on HIV-1 Tat protein. This work would serve as a preliminary test towards HIV detection in real sample.
研究不足
The sensitivity demonstrated in the study is 50 times lower than the reported sensitivity due to the difference in the method used. The study also notes that most GNP-colorimetric assays display sensitivity worse than 5 nM.
1：Experimental Design and Method Selection:
The study utilized an unmodified gold nanoparticle (GNP)-based colorimetric assay for the visible detection of HIV-1 Tat protein. The method involved the use of split aptamers and GNPs to observe color transitions from red to purple in ionic solutions.
2：Sample Selection and Data Sources:
HIV-1 Tat protein was chosen as the target molecule. Split aptamers were purchased from Trilink Biotechnologies, USA, and HIV-1 Tat protein was obtained from Immuno Diagnostics, USA.
3：List of Experimental Equipment and Materials:
Gold nanoparticles (GNPs) of 10 nm size, sodium chloride (NaCl), potassium chloride (KCl), split aptamers, HIV-1 Tat protein, HIV-1 p24, and HIV-1 Nef proteins were used. Instruments included a Lambda 35 UV-Visible Spectrophotometer (Perkin Elmer, USA) and a Supra 35vp Field Emission Scanning Electron Microscope (Zeiss, Germany).
4：Experimental Procedures and Operational Workflow:
The experiment involved titrating GNPs with NaCl to determine the optimum concentration for inducing particle aggregation. Aptamer concentration was optimized to prevent GNP aggregation. Specificity and sensitivity tests were conducted with HIV-1 Tat, p24, and Nef proteins.
5：Data Analysis Methods:
UV-Vis spectrophotometry was used to monitor spectral changes associated with color transitions. FESEM was used to characterize the surface morphology of dispersed and aggregated GNPs.
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