alexa Comparative Evaluation of Molecular Detection Performance of Pseudomonas aeruginosa based on Phylogenetic Markers 16S RNAr, recA, rpoB and ITS1
ISSN: 2327-5073

Clinical Microbiology: Open Access
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Research Article

Comparative Evaluation of Molecular Detection Performance of Pseudomonas aeruginosa based on Phylogenetic Markers 16S RNAr, recA, rpoB and ITS1

Comoé Koffi Donatien Benie1,2*, Adjéhi Dadié1,2, David Coulibaly N’Golo2, Nathalie Guessennd2,3, Solange AKA1,4, Koffi Marcellin DJE1 and Mireille Dosso2,3

1Laboratory of Biotechnology and Food Microbiology (LMBM), University of Nangui-Abrogoua, 02 BP 801 Abidjan 02, Côte d’Ivoire

2Department of Bacteriology and Virology, Institut Pasteur of Côte d'Ivoire (IPCI), 01 BP 490 Abidjan 01, Côte d'Ivoire

3Faculty of Medical Sciences, University of Félix Houphouët Boigny, 01 BP V4 Abidjan 01, Côte d’Ivoire

4Research and Development of Swiss Center for Scientific Research in Côte d'Ivoire, 01 BP 1303 Abidjan 01, Côte d’Ivoire

Corresponding Author:
Comoé Koffi Donatien Benie
Department of Bacteriology and Virology
Institut Pasteur of Côte d'Ivoire (IPCI), 01 BP 490 Abidjan 01, Côte d'Ivoire
Tel: +225 07 55 58 44
E-mail: [email protected]

Received Date: December 4, 2016; Accepted Date: December 22, 2016; Published Date: December 29, 2016

Citation: Benie CKD, Dadié A, N’Golo DC, Guessennd N, Solange AKA, et al. (2016) Comparative Evaluation of Molecular Detection Performance of Pseudomonas aeruginosa based on Phylogenetic Markers 16S RNAr, recA, rpoB and ITS1. Clin Microbiol 5:269. doi: 10.4172/2327-5073.1000269

Copyright: © 2016 Benie CKD, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

 

Abstract

P. aeruginosa may be involved in the poisoning of food. It is highly pathogenic to immunocompromised subjects or weakened, causing a high rate of morbidity and mortality. The aim of this study was to determine the phylogenetic marker suitable for molecular identification of Pseudomonas aeruginosa. The purity and concentration of the nucleic acids were determined by spectrophotometry. Sensitivity reactions using phylogenetic markers (16S RNAr, recA, rpoB, STS1) and the threshold detection of 42 strains were assessed by polymerase chain reaction (PCR). With an average absorption at 230 nm of 2.1, the DNA extracts has an average ratio (A260/A280) of 1.7. The threshold detection of Pseudomonas aeruginosa reference strain ATCC 27853 was 0.8 μg/ml for rpoB and 7.6 μg/ml for each of 16S markers RNAr and recA. The threshold detection of positive control strains CP2: 1125A and CP3: API was 1.2 μg/ml and 0.1 μg/ml for using rpoB gene, respectively. This threshold was respectively 12.3 μg/ml and 0.9 μg/ml for the recA gene. The sensitivity of the rpoB housekeeping gene was 97.4% followed by the recA and 16S RNAr with 87.2% and 82.1%, respectively. The phylogenetic resolution of the rpoB genes was higher than that of the 16S rRNA and recA genes. No sensitivity reaction was observed with ITS1 marker. The quality, purity of the nucleic acids and the choice of phylogenetic marker are among the most critical factors for PCR analysis.

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