Detection of DNA Amplicons of Polymerase Chain Reaction Using Litmus Test

Abstract We report on a new colorimetric DNA detection method that takes advantage of the power of polymerase chain reaction (PCR) and the simplicity of the classic litmus test. The strategy makes use of a modified set of primers for PCR to facilitate ensuing manipulations of resultant DNA amplicons...

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Autores principales:	Dingran Chang, Kha Tram, Ben Li, Qian Feng, Zhifa Shen, Christine H. Lee, Bruno J. Salena, Yingfu Li
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e5f74cf544b74a5a9924eb59a8c265fe
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spelling	oai:doaj.org-article:e5f74cf544b74a5a9924eb59a8c265fe2021-12-02T12:30:12ZDetection of DNA Amplicons of Polymerase Chain Reaction Using Litmus Test10.1038/s41598-017-03009-z2045-2322https://doaj.org/article/e5f74cf544b74a5a9924eb59a8c265fe2017-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-03009-zhttps://doaj.org/toc/2045-2322Abstract We report on a new colorimetric DNA detection method that takes advantage of the power of polymerase chain reaction (PCR) and the simplicity of the classic litmus test. The strategy makes use of a modified set of primers for PCR to facilitate ensuing manipulations of resultant DNA amplicons: their tagging with urease and immobilization onto magnetic beads. The amplicon/urease-laden beads are then used to hydrolyze urea, resulting in the increase of pH that can be conveniently reported by a pH-sensitive dye. We have successfully applied this strategy for the detection of two hypervirulent strains of the bacterium Clostridium difficile that are responsible for the recent increase in the global incidence and severity of C. difficile infections. Furthermore, the viability of this test for diagnostic applications is demonstrated using clinically validated stool samples from C. difficile infected patients.Dingran ChangKha TramBen LiQian FengZhifa ShenChristine H. LeeBruno J. SalenaYingfu LiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-8 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Dingran Chang Kha Tram Ben Li Qian Feng Zhifa Shen Christine H. Lee Bruno J. Salena Yingfu Li Detection of DNA Amplicons of Polymerase Chain Reaction Using Litmus Test
description	Abstract We report on a new colorimetric DNA detection method that takes advantage of the power of polymerase chain reaction (PCR) and the simplicity of the classic litmus test. The strategy makes use of a modified set of primers for PCR to facilitate ensuing manipulations of resultant DNA amplicons: their tagging with urease and immobilization onto magnetic beads. The amplicon/urease-laden beads are then used to hydrolyze urea, resulting in the increase of pH that can be conveniently reported by a pH-sensitive dye. We have successfully applied this strategy for the detection of two hypervirulent strains of the bacterium Clostridium difficile that are responsible for the recent increase in the global incidence and severity of C. difficile infections. Furthermore, the viability of this test for diagnostic applications is demonstrated using clinically validated stool samples from C. difficile infected patients.
format	article
author	Dingran Chang Kha Tram Ben Li Qian Feng Zhifa Shen Christine H. Lee Bruno J. Salena Yingfu Li
author_facet	Dingran Chang Kha Tram Ben Li Qian Feng Zhifa Shen Christine H. Lee Bruno J. Salena Yingfu Li
author_sort	Dingran Chang
title	Detection of DNA Amplicons of Polymerase Chain Reaction Using Litmus Test
title_short	Detection of DNA Amplicons of Polymerase Chain Reaction Using Litmus Test
title_full	Detection of DNA Amplicons of Polymerase Chain Reaction Using Litmus Test
title_fullStr	Detection of DNA Amplicons of Polymerase Chain Reaction Using Litmus Test
title_full_unstemmed	Detection of DNA Amplicons of Polymerase Chain Reaction Using Litmus Test
title_sort	detection of dna amplicons of polymerase chain reaction using litmus test
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/e5f74cf544b74a5a9924eb59a8c265fe
work_keys_str_mv	AT dingranchang detectionofdnaampliconsofpolymerasechainreactionusinglitmustest AT khatram detectionofdnaampliconsofpolymerasechainreactionusinglitmustest AT benli detectionofdnaampliconsofpolymerasechainreactionusinglitmustest AT qianfeng detectionofdnaampliconsofpolymerasechainreactionusinglitmustest AT zhifashen detectionofdnaampliconsofpolymerasechainreactionusinglitmustest AT christinehlee detectionofdnaampliconsofpolymerasechainreactionusinglitmustest AT brunojsalena detectionofdnaampliconsofpolymerasechainreactionusinglitmustest AT yingfuli detectionofdnaampliconsofpolymerasechainreactionusinglitmustest
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Detection of DNA Amplicons of Polymerase Chain Reaction Using Litmus Test

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