Highly Reproducible and Sensitive Electrochemiluminescence Biosensors for HPV Detection Based on Bovine Serum Albumin Carrier Platforms and Hyperbranched Rolling Circle Amplification

• Published in: ACS applied materials & interfaces 2021-01, Vol.13 (1), p.298-305
• Main Authors: He, Yinghao, Liu, Yinhuan, Cheng, Lingjun, Yang, Yuanyuan, Qiu, Bin, Guo, Longhua, Wang, Yan, Lin, Zhenyu, Hong, Guolin
• Format: Article
• Language: English
• Subjects: Animals; Biosensing Techniques - methods; Cattle; Cervix Uteri - virology; DNA Probes - chemistry; DNA Probes - genetics; DNA, Single-Stranded - chemistry; DNA, Single-Stranded - genetics; Electrochemical Techniques - methods; Female; Human papillomavirus 16 - chemistry; Human papillomavirus 16 - isolation & purification; Humans; Limit of Detection; Luminescent Agents; Nucleic Acid Amplification Techniques - methods; Nucleic Acid Hybridization; Oncogene Proteins, Viral - analysis; Oncogene Proteins, Viral - genetics; Organometallic Compounds - chemistry; Papillomavirus E7 Proteins - analysis; Papillomavirus E7 Proteins - genetics; Papillomavirus Infections - diagnosis; Phenanthrolines - chemistry; Repressor Proteins - analysis; Repressor Proteins - genetics; Reproducibility of Results; Ruthenium - chemistry; Serum Albumin, Bovine - chemistry
• ISSN: 1944-8244; 1944-8252
• DOI: 10.1021/acsami.0c20742

Most DNA-based electrochemiluminescence (ECL) biosensors are established through the self-assembly of thiolated single-stranded DNA (ssDNA) probes on the Au electrode surface. Because of this random assembly process, a significant discrepancy exists in the distribution of a modified DNA film on different electrodes, which greatly affects the reproducibility of a biosensor. In this study, a porous bovine serum albumin (BSA) layer was first modified on the electrode surface, which can improve the position distribution and spatial orientation of the self-assembly ssDNA probe. It was then coupled with hyperbranched rolling circle amplification to develop the high-reproducibility-and-sensitivity ECL biosensor for human papillomavirus 16 E6 and E7 oncogene detection. In the presence of the target DNA, the surface of the electrode accumulates abundant amplified products through reaction, which contain double-stranded DNA (dsDNA) fragments of different lengths, followed by plentiful dichlorotris (1,10-phenanthroline) ruthenium(II) hydrate (Ru(phen) , acting as an ECL indicator) insertion into grooves of dsDNA fragments, and a strong signal can be detected. There is a linear relationship between the signal and the target concentration range from 10 fM to 15 pM, and the detection limit is 7.6 fM (S/N = 3). After the BSA modification step, the relative standard deviation was reduced from 9.20 to 3.96%, thereby achieving good reproducibility. The proposed ECL strategy provides a new method for constructing high-reproducibility-and-sensitivity ECL biosensors.