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Transformation of proteins into reproductive DNA templates for sensitive quantification of PSA
Zhao Li1, Jingjing Fu2, Lijuan Wang3
1Key Laboratory of Longevity and Aging-related Diseases of Chinese Ministry of Education, Guangxi Colleges and Universities Key Laboratory of Biological Molecular Medicine Research, School of Basic Medical Sciences, Guangxi Medical University, Nanning, Guangxi, 530021, PR China.
Abstract:
A switch DNA template was designed to transform proteins into linear DNA strands of 97 nt. The linear DNA was subjected to quantitative polymerase chain reaction (qPCR) to determine the initial copy number, which correlated positively with the protein concentration. A restriction endonuclease was used to remove background amplification. Using prostate-specific antigen (PSA) as a protein model, the established method quantified PSA in the range of 10-18-10-13 mol/mL (detection limit = 0.034 pg/mL) with an R2 of 0.974. Good repeatability and specificity of the method were demonstrated. The established method was successfully applied for the quantification of serum PSA levels in women. Significant differences in PSA levels were observed between healthy participants and polycystic ovary syndrome patients.
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