African Journal of
Microbiology Research

  • Abbreviation: Afr. J. Microbiol. Res.
  • Language: English
  • ISSN: 1996-0808
  • DOI: 10.5897/AJMR
  • Start Year: 2007
  • Published Articles: 5232

Full Length Research Paper

Evaluation of real-time PCR for Mycobacterium ulcerans in endemic region in Côte d’Ivoire

Elise Solange Ngazoa-Kakou1,3*, Euloge Ekaza1, Nguetta Aka2, David Coulibaly-N’Golo1, Bakary Coulibaly2 and Mireille Dosso1,2
1Plateforme de Biologie Moléculaire, Institut Pasteur Cote d’Ivoire, BP 490 Abidjan, Cote d’Ivoire. 2Unité des Mycobactéries Tuberculeuses et Atypiques, Institut Pasteur Cote d’Ivoire, BP 490 Abidjan, Cote d’Ivoire. 3Institut des Nutraceutiques et des Aliments Fonctionnels, Université Laval, Qc, G1K 7P4,Canada.
Email: [email protected]

  •  Accepted: 19 May 2011
  •  Published: 18 August 2011

Abstract

Buruli ulcer (BU) is caused by a mycobacterium called Mycobacterium ulcerans. The events of BU are the skin lesions. The lack of early diagnosis and treatment cause severe disability. Today the emergence to BU in Africa and particularly in Côte d’Ivoire needs faster diagnosis to control and to prevent the infection by M. ulcerans. The surveillance of BU is difficult, because the transmission of M. ulcerans occurs in rural regions where the transport of fresh collected sample is long, and the detection with culture technique needs several months. This study has allowed the application of polymerase chain reaction (PCR) technique in real time with two targets for molecular diagnosis of BU in Côte d'Ivoire. 63 samples (clinical, environmental, local strains and reference strains) were analyzed in real-time PCR by comparing the target of the Insertion Sequence (IS) 2404and the sequence Ketoreductase-B (KR-B), located respectively on the chromosome andon the virulence plasmid. 49 samples (76%) were positive in real-time for both targets. The sensitivity of the PCR shows a detection limit of 0.25 genome copy for both targets. The capacity, speed and sensitivity of real-time PCR assays improve the diagnosis and contribute to strengthening the eradication of BU in Côte d’Ivoire.

 

Key words: Buruli ulcer, Mycobacterium ulcerans, real-time, insertion sequence, ketoreductase, Côte d’Ivoire.