Literature DB >> 30269961

Quantification of cDNA on GMR biosensor array towards point-of-care gene expression analysis.

Neeraja Ravi1, Giovanni Rizzi2, Sarah E Chang3, Peggie Cheung4, Paul J Utz5, Shan X Wang6.   

Abstract

Gene expression analysis at the point-of-care is important for rapid disease diagnosis, but traditional techniques are limited by multiplexing capabilities, bulky equipment, and cost. We present a gene expression analysis platform using a giant magnetoresistive (GMR) biosensor array, which allows multiplexed transcript detection and quantification through cost-effective magnetic detection. In this work, we have characterized the sensitivity, dynamic range, and quantification accuracy of Polymerase chain reaction (PCR)-amplified complementary DNA (cDNA) on the GMR for the reference gene GAPDH. A synthetic GAPDH single-stranded DNA (ssDNA) standard was used to calibrate the detection, and ssDNA dilutions were qPCR-amplified to obtain a standard curve. We demonstrate that the GMR platform provides a dynamic range of 4 orders of magnitude and a limit of detection of 1 pM and 0.1 pM respectively for 15 and 18-cycle amplified synthetic GAPDH PCR products. The quantitative results of GMR analysis of cell-line RNA were confirmed by qPCR.
Copyright © 2018 Elsevier B.V. All rights reserved.

Entities:  

Keywords:  Array; DNA melting; GMR sensors; Gene expression; PCR; Point-of-care

Mesh:

Substances:

Year:  2018        PMID: 30269961      PMCID: PMC6391206          DOI: 10.1016/j.bios.2018.09.050

Source DB:  PubMed          Journal:  Biosens Bioelectron        ISSN: 0956-5663            Impact factor:   10.618


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