Coupling Sensitive Nucleic Acid Amplification with Commercial Pregnancy Test Strips

Overview

Journal Angew Chem Int Ed Engl

Specialty Chemistry

Date 2016 Dec 20

PMID 27990727

Citations 29

Authors

Yan Du

Arti Pothukuchy

Jimmy D Gollihar

Armin Nourani

Bingling Li

Andrew D Ellington

Affiliations

Soon will be listed here.

Abstract

The detection of nucleic acid biomarkers for point-of-care (POC) diagnostics is currently limited by technical complexity, cost, and time constraints. To overcome these shortcomings, we have combined loop-mediated isothermal amplification (LAMP), programmable toehold-mediated strand-exchange signal transduction, and standard pregnancy test strips. The incorporation of an engineered hCG-SNAP fusion reporter protein (human chorionic gonadotropin-O -alkylguanine-DNA alkyltransferase) led to LAMP-to-hCG signal transduction on low-cost, commercially available pregnancy test strips. Our assay reliably detected as few as 20 copies of Ebola virus templates in both human serum and saliva and could be adapted to distinguish a common melanoma-associated SNP allele (BRAF V600E) from the wild-type sequence. The methods described are completely generalizable to many nucleic acid biomarkers, and could be adapted to provide POC diagnostics for a range of pathogens.

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References

Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N . Loop-mediated isothermal amplification of DNA. Nucleic Acids Res. 2000; 28(12):E63. PMC: 102748. DOI: 10.1093/nar/28.12.e63. View

Vo-Dinh T, Cullum B . Biosensors and biochips: advances in biological and medical diagnostics. Fresenius J Anal Chem. 2001; 366(6-7):540-51. DOI: 10.1007/s002160051549. View

Nagamine K, Kuzuhara Y, Notomi T . Isolation of single-stranded DNA from loop-mediated isothermal amplification products. Biochem Biophys Res Commun. 2002; 290(4):1195-8. DOI: 10.1006/bbrc.2001.6334. View

Daar A, Thorsteinsdottir H, Martin D, Smith A, Nast S, Singer P . Top ten biotechnologies for improving health in developing countries. Nat Genet. 2002; 32(2):229-32. DOI: 10.1038/ng1002-229. View

Pollock P, Harper U, Hansen K, Yudt L, Stark M, Robbins C . High frequency of BRAF mutations in nevi. Nat Genet. 2002; 33(1):19-20. DOI: 10.1038/ng1054. View

Cheng M, Cuda G, Bunimovich Y, Gaspari M, Heath J, Hill H . Nanotechnologies for biomolecular detection and medical diagnostics. Curr Opin Chem Biol. 2006; 10(1):11-9. DOI: 10.1016/j.cbpa.2006.01.006. View

Wong D . Salivary diagnostics powered by nanotechnologies, proteomics and genomics. J Am Dent Assoc. 2006; 137(3):313-21. DOI: 10.14219/jada.archive.2006.0180. View

Urdea M, Penny L, Olmsted S, Giovanni M, Kaspar P, Shepherd A . Requirements for high impact diagnostics in the developing world. Nature. 2006; 444 Suppl 1:73-9. DOI: 10.1038/nature05448. View

Montagnana M, Caputo M, Giavarina D, Lippi G . Overview on self-monitoring of blood glucose. Clin Chim Acta. 2009; 402(1-2):7-13. DOI: 10.1016/j.cca.2009.01.002. View

10.

Xiang Y, Lu Y . Using personal glucose meters and functional DNA sensors to quantify a variety of analytical targets. Nat Chem. 2011; 3(9):697-703. PMC: 3299819. DOI: 10.1038/nchem.1092. View

11.

Zhang D, Chen S, Yin P . Optimizing the specificity of nucleic acid hybridization. Nat Chem. 2012; 4(3):208-14. PMC: 4238961. DOI: 10.1038/nchem.1246. View

12.

Khunthong S, Jaroenram W, Arunrut N, Suebsing R, Mungsantisuk I, Kiatpathomchai W . Rapid and sensitive detection of shrimp yellow head virus by loop-mediated isothermal amplification combined with a lateral flow dipstick. J Virol Methods. 2012; 188(1-2):51-6. DOI: 10.1016/j.jviromet.2012.11.041. View

13.

Turner A . Biosensors: sense and sensibility. Chem Soc Rev. 2013; 42(8):3184-96. DOI: 10.1039/c3cs35528d. View

14.

Chen S, Zhang D, Seelig G . Conditionally fluorescent molecular probes for detecting single base changes in double-stranded DNA. Nat Chem. 2013; 5(9):782-9. PMC: 3844531. DOI: 10.1038/nchem.1713. View

15.

Zhou Z, Xiang Y, Tong A, Lu Y . Simple and efficient method to purify DNA-protein conjugates and its sensing applications. Anal Chem. 2014; 86(8):3869-75. PMC: 4004194. DOI: 10.1021/ac4040554. View

16.

Yongkiettrakul S, Jaroenram W, Arunrut N, Chareanchim W, Pannengpetch S, Suebsing R . Application of loop-mediated isothermal amplification assay combined with lateral flow dipstick for detection of Plasmodium falciparum and Plasmodium vivax. Parasitol Int. 2014; 63(6):777-84. DOI: 10.1016/j.parint.2014.06.004. View

17.

Zhu Z, Guan Z, Jia S, Lei Z, Lin S, Zhang H . Au@Pt nanoparticle encapsulated target-responsive hydrogel with volumetric bar-chart chip readout for quantitative point-of-care testing. Angew Chem Int Ed Engl. 2014; 53(46):12503-7. DOI: 10.1002/anie.201405995. View

18.

Tram K, Kanda P, Salena B, Huan S, Li Y . Translating bacterial detection by DNAzymes into a litmus test. Angew Chem Int Ed Engl. 2014; 53(47):12799-802. DOI: 10.1002/anie.201407021. View

19.

Jiang Y, Bhadra S, Li B, Wu Y, Milligan J, Ellington A . Robust strand exchange reactions for the sequence-specific, real-time detection of nucleic acid amplicons. Anal Chem. 2015; 87(6):3314-20. DOI: 10.1021/ac504387c. View

20.

Du Y, Hughes R, Bhadra S, Jiang Y, Ellington A, Li B . A Sweet Spot for Molecular Diagnostics: Coupling Isothermal Amplification and Strand Exchange Circuits to Glucometers. Sci Rep. 2015; 5:11039. PMC: 4458886. DOI: 10.1038/srep11039. View