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Biosens Bioelectron. 2019 Jul 1;136:1-7. doi: 10.1016/j.bios.2019.04.027. Epub 2019 Apr 19.

Film-Spotting chiral miniPEG-γPNA array for BRCA1 gene mutation detection.

Author information

1
Hunan Key Laboratory for Super Microstructure and Ultrafast Process, School of Physics and Electronics, Central South University, Changsha, Hunan, 410083, PR China; School of Basic Medical Science, Central South University, Changsha, Hunan, 410083, PR China.
2
Hunan Key Laboratory for Super Microstructure and Ultrafast Process, School of Physics and Electronics, Central South University, Changsha, Hunan, 410083, PR China.
3
State Engineering Laboratory of Highway Maintenance Technology, School of Traffic and Transportation Engineering, Changsha University of Science and Technology, Changsha, 410114, PR China.
4
(d)Xiangya Hospital Central South University, Changsha, 410008, PR China.
5
Hunan Key Laboratory for Super Microstructure and Ultrafast Process, School of Physics and Electronics, Central South University, Changsha, Hunan, 410083, PR China; School of Basic Medical Science, Central South University, Changsha, Hunan, 410083, PR China. Electronic address: liuzhengchunseu@126.com.

Abstract

Peptide nucleic acids array technology is a method of greatly increasing the throughput of laboratory processes to efficiently perform large-scale genetic tests. Diethylene glycol-containing chiral γPNA (miniPEG-γPNA) is considered to be the best PNA derivative and one of the best candidates for gene detection, because it can hybridize DNA with greater affinity and sequence selectivity than DNA and ordinary aminoethylglycyl PNA (aegPNA). Herein, miniPEG-γPNA probes are synthesized by 9-fluorenylmethyloxycarbonyl (Fmoc) solid phase peptide synthesis (SPPS) in a mild condition, and a new biochip fabrication method "Film-Spotting" is invented, by which γPNA arrays with regular pattern, uniform luminance, and very low fluorescence background are obtained easily and cheaply. The miniPEG-γPNA array can effectively distinguish the full matched and mismatched targets in SSarc buffer, serum and urine, and the detection limit of complementary DNA is less than 5.97 nM. A miniPEG-γPNA array for BRCA1 gene mutation (3099delT) detection is also fabricated with a very good detection performance. This work provides an effective avenue for the diagnosis of breast cancer biomarker and expands the application of miniPEG-γPNA in the field of biochip.

KEYWORDS:

BRCA1 mutation; Chiral γPNA; PNA array; Peptide nucleic acid; miniPEG

PMID:
31026759
DOI:
10.1016/j.bios.2019.04.027

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