Effects of decontamination, DNA extraction and amplification procedures on the molecular diagnosis of Mycobacterium ulcerans disease (Buruli ulcer)

D Affolabi; ND Sanoussi; K Vandelannoote; M Odoun; F Faihun; G Sopoh; S Anagonou; F Portaels; M Eddyani

doi:10.1128/JCM.05592-11

Effects of decontamination, DNA extraction and amplification procedures on the molecular diagnosis of Mycobacterium ulcerans disease (Buruli ulcer)

D Affolabi, ND Sanoussi, K Vandelannoote, M Odoun, F Faihun, G Sopoh, S Anagonou, F Portaels, M Eddyani

Mycobacteriology

Research output: Contribution to journal › A1: Web of Science-article › peer-review

37 Downloads (Pure)

Abstract

We compared two DNA extraction methods (a semi-automated method using Maxwell(R) kit and a modified Boom method) and three amplification procedures (a single-step PCR, a nested PCR and a real-time quantitative PCR) on 74 surgical tissue specimens from clinically suspected Buruli ulcer patients. All these procedures were compared before and after decontamination.We observed that, among the procedures tested, real-time PCR after the modified Boom extraction method or a single-run PCR assay after the Maxwell(R) 16 extraction method, performed on non-decontaminated suspensions are the best for the molecular diagnosis of Mycobacterium ulcerans disease.

Original language	English
Journal	Journal of Clinical Microbiology
Volume	50
Issue number	4
Pages (from-to)	1195-1198
ISSN	0095-1137
DOIs	https://doi.org/10.1128/JCM.05592-11
Publication status	Published - 2012

Keywords

B780-tropical-medicine
Bacterial diseases
Buruli ulcer
Mycobacterium ulcerans
Molecular diagnostic techniques
Decontamination
DNA extraction
Amplification
Polymerase chain reaction
PCR
Comparison
Methodology
Smear-positive
Laboratory techniques and procedures

Access to Document

10.1128/JCM.05592-11

2012jcmi1195

http://lib.itg.be/pdf/itg/2012/2012jcmi1195.pdf

Cite this

@article{2ab773ea8bf147519035614b0bb67877,

title = "Effects of decontamination, DNA extraction and amplification procedures on the molecular diagnosis of Mycobacterium ulcerans disease (Buruli ulcer)",

abstract = "We compared two DNA extraction methods (a semi-automated method using Maxwell(R) kit and a modified Boom method) and three amplification procedures (a single-step PCR, a nested PCR and a real-time quantitative PCR) on 74 surgical tissue specimens from clinically suspected Buruli ulcer patients. All these procedures were compared before and after decontamination.We observed that, among the procedures tested, real-time PCR after the modified Boom extraction method or a single-run PCR assay after the Maxwell(R) 16 extraction method, performed on non-decontaminated suspensions are the best for the molecular diagnosis of Mycobacterium ulcerans disease.",

keywords = "B780-tropical-medicine, Bacterial diseases, Buruli ulcer, Mycobacterium ulcerans, Molecular diagnostic techniques, Decontamination, DNA extraction, Amplification, Polymerase chain reaction, PCR, Comparison, Methodology, Smear-positive, Laboratory techniques and procedures",

author = "D Affolabi and ND Sanoussi and K Vandelannoote and M Odoun and F Faihun and G Sopoh and S Anagonou and F Portaels and M Eddyani",

note = "FTX; ITG-B3B; ITG-X8B; ITG-BLB; DBM; U-MYCOB; JIF; DOI; PDF; Abstract; DSPACE",

year = "2012",

doi = "10.1128/JCM.05592-11",

language = "English",

volume = "50",

pages = "1195--1198",

journal = "Journal of Clinical Microbiology",

issn = "0095-1137",

publisher = "American Society for Microbiology",

number = "4",

}

Affolabi, D, Sanoussi, ND, Vandelannoote, K, Odoun, M, Faihun, F, Sopoh, G, Anagonou, S, Portaels, F & Eddyani, M 2012, 'Effects of decontamination, DNA extraction and amplification procedures on the molecular diagnosis of Mycobacterium ulcerans disease (Buruli ulcer)', Journal of Clinical Microbiology, vol. 50, no. 4, pp. 1195-1198. https://doi.org/10.1128/JCM.05592-11

TY - JOUR

T1 - Effects of decontamination, DNA extraction and amplification procedures on the molecular diagnosis of Mycobacterium ulcerans disease (Buruli ulcer)

AU - Affolabi, D

AU - Sanoussi, ND

AU - Vandelannoote, K

AU - Odoun, M

AU - Faihun, F

AU - Sopoh, G

AU - Anagonou, S

AU - Portaels, F

AU - Eddyani, M

N1 - FTX; ITG-B3B; ITG-X8B; ITG-BLB; DBM; U-MYCOB; JIF; DOI; PDF; Abstract; DSPACE

PY - 2012

Y1 - 2012

N2 - We compared two DNA extraction methods (a semi-automated method using Maxwell(R) kit and a modified Boom method) and three amplification procedures (a single-step PCR, a nested PCR and a real-time quantitative PCR) on 74 surgical tissue specimens from clinically suspected Buruli ulcer patients. All these procedures were compared before and after decontamination.We observed that, among the procedures tested, real-time PCR after the modified Boom extraction method or a single-run PCR assay after the Maxwell(R) 16 extraction method, performed on non-decontaminated suspensions are the best for the molecular diagnosis of Mycobacterium ulcerans disease.

AB - We compared two DNA extraction methods (a semi-automated method using Maxwell(R) kit and a modified Boom method) and three amplification procedures (a single-step PCR, a nested PCR and a real-time quantitative PCR) on 74 surgical tissue specimens from clinically suspected Buruli ulcer patients. All these procedures were compared before and after decontamination.We observed that, among the procedures tested, real-time PCR after the modified Boom extraction method or a single-run PCR assay after the Maxwell(R) 16 extraction method, performed on non-decontaminated suspensions are the best for the molecular diagnosis of Mycobacterium ulcerans disease.

KW - B780-tropical-medicine

KW - Bacterial diseases

KW - Buruli ulcer

KW - Mycobacterium ulcerans

KW - Molecular diagnostic techniques

KW - Decontamination

KW - DNA extraction

KW - Amplification

KW - Polymerase chain reaction

KW - PCR

KW - Comparison

KW - Methodology

KW - Smear-positive

KW - Laboratory techniques and procedures

U2 - 10.1128/JCM.05592-11

DO - 10.1128/JCM.05592-11

M3 - A1: Web of Science-article

C2 - 22259213

SN - 0095-1137

VL - 50

SP - 1195

EP - 1198

JO - Journal of Clinical Microbiology

JF - Journal of Clinical Microbiology

IS - 4

ER -

Effects of decontamination, DNA extraction and amplification procedures on the molecular diagnosis of Mycobacterium ulcerans disease (Buruli ulcer)

Abstract

Keywords

Access to Document

Fingerprint

Cite this