Molecular Detection of <i>Borrelia</i> Bacteria in Cerebrospinal Fluid-Optimisation of Pre-Analytical Sample Handling for Increased Analytical Sensitivity

The main tools for clinical diagnostics of Lyme neuroborreliosis (LNB) are based on serology, i.e., detection of antibodies in cerebrospinal fluid (CSF). In some cases, PCR may be used as a supplement, e.g., on CSF from patients with early LNB. Standardisation of the molecular methods and systematic...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Malin Lager, Peter Wilhelmsson, Andreas Matussek, Per-Eric Lindgren, Anna J. Henningsson
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	Lyme borreliosis Lyme neuroborreliosis polymerase chain reaction (PCR) cerebrospinal fluid clinical diagnostic optimization Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/b38cf3da9abf4f79a5d27e6bac2b3b00
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:b38cf3da9abf4f79a5d27e6bac2b3b00
record_format	dspace
spelling	oai:doaj.org-article:b38cf3da9abf4f79a5d27e6bac2b3b002021-11-25T17:21:31ZMolecular Detection of <i>Borrelia</i> Bacteria in Cerebrospinal Fluid-Optimisation of Pre-Analytical Sample Handling for Increased Analytical Sensitivity10.3390/diagnostics111120882075-4418https://doaj.org/article/b38cf3da9abf4f79a5d27e6bac2b3b002021-11-01T00:00:00Zhttps://www.mdpi.com/2075-4418/11/11/2088https://doaj.org/toc/2075-4418The main tools for clinical diagnostics of Lyme neuroborreliosis (LNB) are based on serology, i.e., detection of antibodies in cerebrospinal fluid (CSF). In some cases, PCR may be used as a supplement, e.g., on CSF from patients with early LNB. Standardisation of the molecular methods and systematic evaluation of the pre-analytical handling is lacking. To increase the analytical sensitivity for detection of <i>Borrelia</i> bacteria in CSF by PCR targeting the <i>16S</i> rRNA gene, parameters were systematically evaluated on CSF samples spiked with a known amount of cultured <i>Borrelia</i> bacteria. The results showed that the parameters such as centrifugation time and speed, the use of complementary DNA as a template (in combination with primers and a probe aiming at target gene <i>16S</i> rRNA), and the absence of inhibitors (e.g., erythrocytes) had the highest impact on the analytical sensitivity. Based on these results, a protocol for optimised handling of CSF samples before molecular analysis was proposed. However, no clinical evaluation of the proposed protocol has been done so far, and further investigations of the diagnostic sensitivity need to be performed on well-characterised clinical samples from patients with LNB.Malin LagerPeter WilhelmssonAndreas MatussekPer-Eric LindgrenAnna J. HenningssonMDPI AGarticleLyme borreliosisLyme neuroborreliosispolymerase chain reaction (PCR)cerebrospinal fluidclinical diagnosticoptimizationMedicine (General)R5-920ENDiagnostics, Vol 11, Iss 2088, p 2088 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Lyme borreliosis Lyme neuroborreliosis polymerase chain reaction (PCR) cerebrospinal fluid clinical diagnostic optimization Medicine (General) R5-920
spellingShingle	Lyme borreliosis Lyme neuroborreliosis polymerase chain reaction (PCR) cerebrospinal fluid clinical diagnostic optimization Medicine (General) R5-920 Malin Lager Peter Wilhelmsson Andreas Matussek Per-Eric Lindgren Anna J. Henningsson Molecular Detection of <i>Borrelia</i> Bacteria in Cerebrospinal Fluid-Optimisation of Pre-Analytical Sample Handling for Increased Analytical Sensitivity
description	The main tools for clinical diagnostics of Lyme neuroborreliosis (LNB) are based on serology, i.e., detection of antibodies in cerebrospinal fluid (CSF). In some cases, PCR may be used as a supplement, e.g., on CSF from patients with early LNB. Standardisation of the molecular methods and systematic evaluation of the pre-analytical handling is lacking. To increase the analytical sensitivity for detection of <i>Borrelia</i> bacteria in CSF by PCR targeting the <i>16S</i> rRNA gene, parameters were systematically evaluated on CSF samples spiked with a known amount of cultured <i>Borrelia</i> bacteria. The results showed that the parameters such as centrifugation time and speed, the use of complementary DNA as a template (in combination with primers and a probe aiming at target gene <i>16S</i> rRNA), and the absence of inhibitors (e.g., erythrocytes) had the highest impact on the analytical sensitivity. Based on these results, a protocol for optimised handling of CSF samples before molecular analysis was proposed. However, no clinical evaluation of the proposed protocol has been done so far, and further investigations of the diagnostic sensitivity need to be performed on well-characterised clinical samples from patients with LNB.
format	article
author	Malin Lager Peter Wilhelmsson Andreas Matussek Per-Eric Lindgren Anna J. Henningsson
author_facet	Malin Lager Peter Wilhelmsson Andreas Matussek Per-Eric Lindgren Anna J. Henningsson
author_sort	Malin Lager
title	Molecular Detection of <i>Borrelia</i> Bacteria in Cerebrospinal Fluid-Optimisation of Pre-Analytical Sample Handling for Increased Analytical Sensitivity
title_short	Molecular Detection of <i>Borrelia</i> Bacteria in Cerebrospinal Fluid-Optimisation of Pre-Analytical Sample Handling for Increased Analytical Sensitivity
title_full	Molecular Detection of <i>Borrelia</i> Bacteria in Cerebrospinal Fluid-Optimisation of Pre-Analytical Sample Handling for Increased Analytical Sensitivity
title_fullStr	Molecular Detection of <i>Borrelia</i> Bacteria in Cerebrospinal Fluid-Optimisation of Pre-Analytical Sample Handling for Increased Analytical Sensitivity
title_full_unstemmed	Molecular Detection of <i>Borrelia</i> Bacteria in Cerebrospinal Fluid-Optimisation of Pre-Analytical Sample Handling for Increased Analytical Sensitivity
title_sort	molecular detection of <i>borrelia</i> bacteria in cerebrospinal fluid-optimisation of pre-analytical sample handling for increased analytical sensitivity
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/b38cf3da9abf4f79a5d27e6bac2b3b00
work_keys_str_mv	AT malinlager moleculardetectionofiborreliaibacteriaincerebrospinalfluidoptimisationofpreanalyticalsamplehandlingforincreasedanalyticalsensitivity AT peterwilhelmsson moleculardetectionofiborreliaibacteriaincerebrospinalfluidoptimisationofpreanalyticalsamplehandlingforincreasedanalyticalsensitivity AT andreasmatussek moleculardetectionofiborreliaibacteriaincerebrospinalfluidoptimisationofpreanalyticalsamplehandlingforincreasedanalyticalsensitivity AT perericlindgren moleculardetectionofiborreliaibacteriaincerebrospinalfluidoptimisationofpreanalyticalsamplehandlingforincreasedanalyticalsensitivity AT annajhenningsson moleculardetectionofiborreliaibacteriaincerebrospinalfluidoptimisationofpreanalyticalsamplehandlingforincreasedanalyticalsensitivity
_version_	1718412492137299968

Molecular Detection of <i>Borrelia</i> Bacteria in Cerebrospinal Fluid-Optimisation of Pre-Analytical Sample Handling for Increased Analytical Sensitivity

Ejemplares similares