Aarhus University Seal / Aarhus Universitets segl

A new DNA sensor system for specific and quantitative detection of mycobacteria

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

Standard

A new DNA sensor system for specific and quantitative detection of mycobacteria. / Franch, Oskar; Han, Xiao; Marcussen, Lærke; Jørgensen, Asger Givskov; Andersen, Marie Bech; Godbole, Adwait Anand; Harmsen, Charlotte; Nørskov-Lauritsen, Niels; Thomsen, Jonas; Pedersen, Finn Skou; Wang, Yilong; Shi, Donglu; Wejse, Christian; Pødenphant, Lone; Nagaraja, Valakunja; Bertl, Johanna; Stougaard, Magnus; Ho, Yi-Ping; Hede, Marianne Smedegaard; Labouriau, Rodrigo; Knudsen, Birgitta Ruth.

I: Nanoscale, Bind 11, Nr. 2, 14.01.2019, s. 587-597.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

Harvard

APA

CBE

MLA

Vancouver

Author

Franch, Oskar ; Han, Xiao ; Marcussen, Lærke ; Jørgensen, Asger Givskov ; Andersen, Marie Bech ; Godbole, Adwait Anand ; Harmsen, Charlotte ; Nørskov-Lauritsen, Niels ; Thomsen, Jonas ; Pedersen, Finn Skou ; Wang, Yilong ; Shi, Donglu ; Wejse, Christian ; Pødenphant, Lone ; Nagaraja, Valakunja ; Bertl, Johanna ; Stougaard, Magnus ; Ho, Yi-Ping ; Hede, Marianne Smedegaard ; Labouriau, Rodrigo ; Knudsen, Birgitta Ruth. / A new DNA sensor system for specific and quantitative detection of mycobacteria. I: Nanoscale. 2019 ; Bind 11, Nr. 2. s. 587-597.

Bibtex

@article{ff8a6e5fa5f2467094873cc9e2f6206e,
title = "A new DNA sensor system for specific and quantitative detection of mycobacteria",
abstract = "In the current study, we describe a novel DNA sensor system for specific and quantitative detection of mycobacteria, which is the causative agent of tuberculosis. Detection is achieved by using the enzymatic activity of the mycobacterial encoded enzyme topoisomerase IA (TOP1A) as a biomarker. The presented work is the first to describe how the catalytic activities of a member of the type IA family of topoisomerases can be exploited for specific detection of bacteria. The principle for detection relies on a solid support anchored DNA substrate with dual functions namely: (1) the ability to isolate mycobacterial TOP1A from crude samples and (2) the ability to be converted into a closed DNA circle upon reaction with the isolated enzyme. The DNA circle can act as a template for rolling circle amplification generating a tandem repeat product that can be visualized at the single molecule level by fluorescent labelling. This reaction scheme ensures specific, sensitive, and quantitative detection of the mycobacteria TOP1A biomarker as demonstrated by the use of purified mycobacterial TOP1A and extracts from an array of non-mycobacteria and mycobacteria species. When combined with mycobacteriophage induced lysis as a novel way of effective yet gentle extraction of the cellular content from the model Mycobacterium smegmatis, the DNA sensor system allowed detection of mycobacteria in small volumes of cell suspensions. Moreover, it was possible to detect M. smegmatis added to human saliva. Depending on the composition of the sample, we were able to detect 0.6 or 0.9 million colony forming units (CFU) per mL of mycobacteria, which is within the range of clinically relevant infection numbers. We, therefore, believe that the presented assay, which relies on techniques that can be adapted to limited resource settings, may be the first step towards the development of a new point-of-care diagnostic test for tuberculosis.",
author = "Oskar Franch and Xiao Han and L{\ae}rke Marcussen and J{\o}rgensen, {Asger Givskov} and Andersen, {Marie Bech} and Godbole, {Adwait Anand} and Charlotte Harmsen and Niels N{\o}rskov-Lauritsen and Jonas Thomsen and Pedersen, {Finn Skou} and Yilong Wang and Donglu Shi and Christian Wejse and Lone P{\o}denphant and Valakunja Nagaraja and Johanna Bertl and Magnus Stougaard and Yi-Ping Ho and Hede, {Marianne Smedegaard} and Rodrigo Labouriau and Knudsen, {Birgitta Ruth}",
year = "2019",
month = jan,
day = "14",
doi = "10.1039/c8nr07850e",
language = "English",
volume = "11",
pages = "587--597",
journal = "Nanoscale",
issn = "2040-3364",
publisher = "ROYAL SOC CHEMISTRY",
number = "2",

}

RIS

TY - JOUR

T1 - A new DNA sensor system for specific and quantitative detection of mycobacteria

AU - Franch, Oskar

AU - Han, Xiao

AU - Marcussen, Lærke

AU - Jørgensen, Asger Givskov

AU - Andersen, Marie Bech

AU - Godbole, Adwait Anand

AU - Harmsen, Charlotte

AU - Nørskov-Lauritsen, Niels

AU - Thomsen, Jonas

AU - Pedersen, Finn Skou

AU - Wang, Yilong

AU - Shi, Donglu

AU - Wejse, Christian

AU - Pødenphant, Lone

AU - Nagaraja, Valakunja

AU - Bertl, Johanna

AU - Stougaard, Magnus

AU - Ho, Yi-Ping

AU - Hede, Marianne Smedegaard

AU - Labouriau, Rodrigo

AU - Knudsen, Birgitta Ruth

PY - 2019/1/14

Y1 - 2019/1/14

N2 - In the current study, we describe a novel DNA sensor system for specific and quantitative detection of mycobacteria, which is the causative agent of tuberculosis. Detection is achieved by using the enzymatic activity of the mycobacterial encoded enzyme topoisomerase IA (TOP1A) as a biomarker. The presented work is the first to describe how the catalytic activities of a member of the type IA family of topoisomerases can be exploited for specific detection of bacteria. The principle for detection relies on a solid support anchored DNA substrate with dual functions namely: (1) the ability to isolate mycobacterial TOP1A from crude samples and (2) the ability to be converted into a closed DNA circle upon reaction with the isolated enzyme. The DNA circle can act as a template for rolling circle amplification generating a tandem repeat product that can be visualized at the single molecule level by fluorescent labelling. This reaction scheme ensures specific, sensitive, and quantitative detection of the mycobacteria TOP1A biomarker as demonstrated by the use of purified mycobacterial TOP1A and extracts from an array of non-mycobacteria and mycobacteria species. When combined with mycobacteriophage induced lysis as a novel way of effective yet gentle extraction of the cellular content from the model Mycobacterium smegmatis, the DNA sensor system allowed detection of mycobacteria in small volumes of cell suspensions. Moreover, it was possible to detect M. smegmatis added to human saliva. Depending on the composition of the sample, we were able to detect 0.6 or 0.9 million colony forming units (CFU) per mL of mycobacteria, which is within the range of clinically relevant infection numbers. We, therefore, believe that the presented assay, which relies on techniques that can be adapted to limited resource settings, may be the first step towards the development of a new point-of-care diagnostic test for tuberculosis.

AB - In the current study, we describe a novel DNA sensor system for specific and quantitative detection of mycobacteria, which is the causative agent of tuberculosis. Detection is achieved by using the enzymatic activity of the mycobacterial encoded enzyme topoisomerase IA (TOP1A) as a biomarker. The presented work is the first to describe how the catalytic activities of a member of the type IA family of topoisomerases can be exploited for specific detection of bacteria. The principle for detection relies on a solid support anchored DNA substrate with dual functions namely: (1) the ability to isolate mycobacterial TOP1A from crude samples and (2) the ability to be converted into a closed DNA circle upon reaction with the isolated enzyme. The DNA circle can act as a template for rolling circle amplification generating a tandem repeat product that can be visualized at the single molecule level by fluorescent labelling. This reaction scheme ensures specific, sensitive, and quantitative detection of the mycobacteria TOP1A biomarker as demonstrated by the use of purified mycobacterial TOP1A and extracts from an array of non-mycobacteria and mycobacteria species. When combined with mycobacteriophage induced lysis as a novel way of effective yet gentle extraction of the cellular content from the model Mycobacterium smegmatis, the DNA sensor system allowed detection of mycobacteria in small volumes of cell suspensions. Moreover, it was possible to detect M. smegmatis added to human saliva. Depending on the composition of the sample, we were able to detect 0.6 or 0.9 million colony forming units (CFU) per mL of mycobacteria, which is within the range of clinically relevant infection numbers. We, therefore, believe that the presented assay, which relies on techniques that can be adapted to limited resource settings, may be the first step towards the development of a new point-of-care diagnostic test for tuberculosis.

U2 - 10.1039/c8nr07850e

DO - 10.1039/c8nr07850e

M3 - Journal article

C2 - 30556557

VL - 11

SP - 587

EP - 597

JO - Nanoscale

JF - Nanoscale

SN - 2040-3364

IS - 2

ER -