Copy Number Variation Analysis by Ligation-Dependent PCR Based on Magnetic Nanoparticles and Chemiluminescence

Abstract

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.

Multifunctional Device for Nucleic Acid Extraction Based on Magnetic Separation and Its Co-Working with Liquid Handling System for High Throughput Sample Preparation

“…The application of magnetic nanoparticles for DNA and RNA isolation, also known Copyright: American Scientific Publishers as magnetic nucleic acids separation technology, has been widely accepted as a reliable method. [23][24][25][26][27][28][29][30][31][32] Automations based on this method had gained much attention. 17 20 33-35 However, those existing robotic solutions based on this technology are still far from perfect.…”

Chelerythrine and Fe3O4 Loaded Multi-Walled Carbon Nanotubes for Targeted Cancer Therapy

“…Functionalization of the CNTs with magnetic Fe 3 O 4 nanoparticles (NPs) has been suggested to be very important in developing the targeting ability of the oxidized-CNTs. The multifunctional properties of the Fe 3 O 4 NPs, such as small size, high magnetic responsivity and low toxicity, [34][35][36][37][38][39][40][41][42][43] can be used to increase the accumulation of drugs in pathological sites, thereby reducing the non-specific toxicity and some other adverse side-effects. We therefore decorated the MWNTs with Fe 3 O 4 NPs to ensure a strong magnetic responsiveness and provide the targeted property for various applications.…”

Development and Validation of an Automated Liquid Handling System for Sample Preparation Based on Multichannel Air Displacement Pipetting Technology

“…[7][8][9][10][11][12] High-throughput liquid handling technology is highly needed in life science laboratories because of its ability to process samples rapidly and tirelessly, especially in those integrated devices or instruments for high-throughput bioassays with the help of nanotechnology. [13][14][15][16][17][18][19][20][21][22][23][24][25][26] With advances in nanotechnology, [27][28][29][30][31][32][33][34][35][36][37] particularly in magnetic * Authors to whom correspondence should be addressed. nanotechnology, [38][39][40][41][42][43][44][45][46][47][48][49] high-throughput liquid handling technology can greatly increase the throughput of a laboratory and save precious time for scientists.…”