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Kahla I.,Laboratoire Of Microbiologie Et Dimmunologie | Kahla I.,Aix - Marseille University | Henry M.,Aix - Marseille University | Boukadida J.,Laboratoire Of Microbiologie Et Dimmunologie | Drancourt M.,Aix - Marseille University
BMC Research Notes | Year: 2011

Abstract. Background: Identification of the Mycobacterium tuberculosis complex organisms to the species level is important for diagnostic, therapeutic and epidemiologic perspectives. Indeed, isolates are routinely identified as belonging to the M. tuberculosis complex without further discrimination in agreement with the high genomic similarity of the M. tuberculosis complex members and the resulting complex available identification tools. Findings. We herein develop a pyrosequencing assay analyzing polymorphisms within glpK, pykA and gyrB genes to identify members of the M. tuberculosis complex at the species level. The assay was evaluated with 22 M. tuberculosis, 21 M. bovis, 3 M. caprae, 3 M. microti, 2 M. bovis BCG, 2 M. pinnipedii, 1 M. canettii and 1 M. africanum type I isolates. The resulted pyrograms were consistent with conventional DNA sequencing data and successfully identified all isolates. Additionally, 127 clinical M. tuberculosis complex isolates were analyzed and were unambiguously identified as M. tuberculosis. Conclusion: We proposed a pyrosequencing-based scheme for the rapid identification of M. tuberculosis complex isolates at the species level. The assay is robust, specific, rapid and can be easily introduced in the routine activity. © 2011 Drancourt et al; licensee BioMed Central Ltd.

Ben Kahla I.,Laboratoire Of Microbiologie Et Dimmunologie | Ben Kahla I.,Aix - Marseille University | Marzouk M.,Laboratoire Of Microbiologie Et Dimmunologie | Henry M.,Aix - Marseille University | And 5 more authors.
International Journal of Tuberculosis and Lung Disease | Year: 2011

The aim of our study was to genotypically characterise isoniazid (INH) and rifampicin (RMP) resistant Mycobacterium tuberculosis isolates in Sousse, Central Tunisia, using DNA sequencing and multispacer sequence typing (MST). The results show that 27/28 (96.4%) and 1/28 (3.6%) INH-resistant isolates yielded respectively the katG S315T and the inhA -15C→ mutations. Two-thirds of RMP-resistant isolates yielded the rpoB D516V mutation and one sixth yielded either H526D or S531L mutations. Genotyping analysis revealed the multiclonal spread of drug-resistant isolates in Central Tunisia. Data presented here complete the previously published map of resistant M. tuberculosis isolates and highlight their regional disparity in Tunisia. © 2011 The Union.

Ferjani A.,Laboratoire Of Microbiologie Et Dimmunologie | Marzouk M.,Laboratoire Of Microbiologie Et Dimmunologie | Idriss N.,Laboratoire Of Microbiologie Et Dimmunologie | Sammoud S.,Laboratoire Of Microbiologie Et Dimmunologie | And 2 more authors.
Annales de Biologie Clinique | Year: 2011

We evaluated the performance and the cost of chromogenic medium Uriselect4 agar with regard to the standard medium for the detection and identification of urinary tract pathogens. A total of 503 clinical urine specimens containing leucocytes greater or equal to 10 4/mL were analysed prospectively, in parallel by two different persons on blood agar (GS) and Uriselect4 according to the manufacturers' instructions. Of the 503 urine specimens tested, 210 gave a positive culture on Uriselect4 versus 181 on GS. The majority of bacterial species grew on both media; enterobacteria grew on Uriselect4 better than GS. The identification of Escherichia coli (E. coli), Proteus mirabilis (P. mirabilis), KES group and Enterococcus faecalis (E. faecalis) did not require the use of galleries Api and has a gain of 24 h. Positive pure cultures on Uriselect4 corresponding to negative cultures of GS were noted in 17 cases. Conversely, in seven cases a positive pure culture on GS was noted while the corresponding Uriselect4 cultures were negative. The cost of identification on GS (including the cost of galleries Api), was about two times higher than Uriselect4. Uriselect4 medium isolates the most frequent urinary tract pathogens and identify them so almost immediately, with a lower cost.

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