<i>LoopTag</i> FRET Probe System for Multiplex qPCR Detection of <i>Borrelia</i> Species

Background: Laboratory diagnosis of Lyme borreliosis refers to some methods with known limitations. Molecular diagnostics using specific nucleic acid probes may overcome some of these limitations. Methods: We describe the novel reporter fluorescence real-time polymerase chain reaction (PCR) probe sy...

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Autores principales:	Henning Hanschmann, Stefan Rödiger, Toni Kramer, Katrin Hanschmann, Michael Steidle, Volker Fingerle, Carsten Schmidt, Werner Lehmann, Peter Schierack
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	<i>Borrelia</i> <i>B. Burgdorferi</i> (sensu lato) complex real-time PCR Lyme borreliosis diagnostic Science Q
Acceso en línea:	https://doaj.org/article/a4d9894e284b430d8a85d72e2c4f4371
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spelling	oai:doaj.org-article:a4d9894e284b430d8a85d72e2c4f43712021-11-25T18:10:49Z<i>LoopTag</i> FRET Probe System for Multiplex qPCR Detection of <i>Borrelia</i> Species10.3390/life111111632075-1729https://doaj.org/article/a4d9894e284b430d8a85d72e2c4f43712021-10-01T00:00:00Zhttps://www.mdpi.com/2075-1729/11/11/1163https://doaj.org/toc/2075-1729Background: Laboratory diagnosis of Lyme borreliosis refers to some methods with known limitations. Molecular diagnostics using specific nucleic acid probes may overcome some of these limitations. Methods: We describe the novel reporter fluorescence real-time polymerase chain reaction (PCR) probe system <i>LoopTag</i> for detection of <i>Borrelia</i> species. Advantages of the <i>LoopTag</i> system include having cheap conventional fluorescence dyes, easy primer design, no restrictions for PCR product lengths, robustness, high sequence specificity, applicability for multiplex real-time PCRs, melting curve analysis (single nucleotide polymorphism analysis) over a large temperature range, high sensitivity, and easy adaptation of conventional PCRs. Results: Using the <i>LoopTag</i> probe system we were able to detect all nine tested European species belonging to the <i>Borrelia burgdorferi</i> (sensu lato) complex and differentiated them from relapsing fever <i>Borrelia</i> species. As few as 10 copies of <i>Borrelia</i> in one PCR reaction were detectable. Conclusion: We established a novel multiplex probe real-time PCR system, designated <i>LoopTag</i>, that is simple, robust, and incorporates melting curve analysis for the detection and in the differentiation of European species belonging to the <i>Borrelia burgdorferi</i> s.l. complex.Henning HanschmannStefan RödigerToni KramerKatrin HanschmannMichael SteidleVolker FingerleCarsten SchmidtWerner LehmannPeter SchierackMDPI AGarticle<i>Borrelia</i><i>B. Burgdorferi</i> (sensu lato) complexreal-time PCRLyme borreliosisdiagnosticScienceQENLife, Vol 11, Iss 1163, p 1163 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	<i>Borrelia</i> <i>B. Burgdorferi</i> (sensu lato) complex real-time PCR Lyme borreliosis diagnostic Science Q
spellingShingle	<i>Borrelia</i> <i>B. Burgdorferi</i> (sensu lato) complex real-time PCR Lyme borreliosis diagnostic Science Q Henning Hanschmann Stefan Rödiger Toni Kramer Katrin Hanschmann Michael Steidle Volker Fingerle Carsten Schmidt Werner Lehmann Peter Schierack <i>LoopTag</i> FRET Probe System for Multiplex qPCR Detection of <i>Borrelia</i> Species
description	Background: Laboratory diagnosis of Lyme borreliosis refers to some methods with known limitations. Molecular diagnostics using specific nucleic acid probes may overcome some of these limitations. Methods: We describe the novel reporter fluorescence real-time polymerase chain reaction (PCR) probe system <i>LoopTag</i> for detection of <i>Borrelia</i> species. Advantages of the <i>LoopTag</i> system include having cheap conventional fluorescence dyes, easy primer design, no restrictions for PCR product lengths, robustness, high sequence specificity, applicability for multiplex real-time PCRs, melting curve analysis (single nucleotide polymorphism analysis) over a large temperature range, high sensitivity, and easy adaptation of conventional PCRs. Results: Using the <i>LoopTag</i> probe system we were able to detect all nine tested European species belonging to the <i>Borrelia burgdorferi</i> (sensu lato) complex and differentiated them from relapsing fever <i>Borrelia</i> species. As few as 10 copies of <i>Borrelia</i> in one PCR reaction were detectable. Conclusion: We established a novel multiplex probe real-time PCR system, designated <i>LoopTag</i>, that is simple, robust, and incorporates melting curve analysis for the detection and in the differentiation of European species belonging to the <i>Borrelia burgdorferi</i> s.l. complex.
format	article
author	Henning Hanschmann Stefan Rödiger Toni Kramer Katrin Hanschmann Michael Steidle Volker Fingerle Carsten Schmidt Werner Lehmann Peter Schierack
author_facet	Henning Hanschmann Stefan Rödiger Toni Kramer Katrin Hanschmann Michael Steidle Volker Fingerle Carsten Schmidt Werner Lehmann Peter Schierack
author_sort	Henning Hanschmann
title	<i>LoopTag</i> FRET Probe System for Multiplex qPCR Detection of <i>Borrelia</i> Species
title_short	<i>LoopTag</i> FRET Probe System for Multiplex qPCR Detection of <i>Borrelia</i> Species
title_full	<i>LoopTag</i> FRET Probe System for Multiplex qPCR Detection of <i>Borrelia</i> Species
title_fullStr	<i>LoopTag</i> FRET Probe System for Multiplex qPCR Detection of <i>Borrelia</i> Species
title_full_unstemmed	<i>LoopTag</i> FRET Probe System for Multiplex qPCR Detection of <i>Borrelia</i> Species
title_sort	<i>looptag</i> fret probe system for multiplex qpcr detection of <i>borrelia</i> species
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/a4d9894e284b430d8a85d72e2c4f4371
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<i>LoopTag</i> FRET Probe System for Multiplex qPCR Detection of <i>Borrelia</i> Species

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