Journal
THERANOSTICS
Volume 5, Issue 1, Pages 71-85Publisher
IVYSPRING INT PUBL
DOI: 10.7150/thno.10117
Keywords
Copy number variation; Ligation-dependent PCR; Magnetic nanoparticles; Chemiluminescence
Categories
Funding
- National Basic Research Program (973 Program) of China [2010CB933903, 2014CB744501]
- National Nature Science Foundation of China (NSFC) [61271056, 31201003, 81301305, 21205013, 61471168]
- state key laboratory of bioelectronics of southeast university [2012F12, 2012F13]
- Natural Science Foundation of Jiangsu Province [BK20130892]
Ask authors/readers for more resources
A novel system for copy number variation (CNV) analysis was developed in the present study using a combination of magnetic separation and chemiluminescence (CL) detection technique. The amino-modified probes were firstly immobilized onto carboxylated magnetic nanoparticles (MNPs) and then hybridized with biotin-dUTP products, followed by amplification with ligation-dependent polymerase chain reaction (PCR). After streptavidin-modified alkaline phosphatase (STV-AP) bonding and magnetic separation, the CL signals were then detected. Results showed that the quantification of PCR products could be reflected by CL signal values. Under optimum conditions, the CL system was characterized for quantitative analysis and the CL intensity exhibited a linear correlation with logarithm of the target concentration. To validate the methodology, copy numbers of six genes from the human genome were detected. To compare the detection accuracy, multiplex ligation-dependent probe amplification (MLPA) and MNPs-CL detection were performed. Overall, there were two discrepancies by MLPA analysis, while only one by MNPs-CL detection. This research demonstrated that the novel MNPs-CL system is a useful analytical tool which shows simple, sensitive, and specific characters which are suitable for CNV analysis. Moreover, this system should be improved further and its application in the genome variation detection of various diseases is currently under further investigation.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available