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Mouttaki T.,Center for Doctoral Studies in Health science | Mouttaki T.,Research Team on Cutaneous Leishmaniasis | Chiheb S.,University Hospital Ibn Rochd | Chiheb S.,Research Team on Cutaneous Leishmaniasis | And 4 more authors.
Parasites and Vectors | Year: 2014

Background: The diagnosis of cutaneous leishmaniasis (CL) might be difficult, in particular in endemic areas where different species of Leishmania can cause lesions of very similar appearance and where other skin diseases with similar clinical symptoms occur. Even today, the parasitological diagnosis of CL remains the gold standard and it is based on the direct identification of amastigotes in microscopy smears and/or culture of promastigotes from infected tissues. Although these techniques are highly specific, they are not sensitive enough. The objective of this study is to contribute to improving the diagnosis of CL and the identification of Leishmania species in Morocco by comparing three PCR-based assays applied directly on dermal samples.Methods: A total of 58 patients presenting with cutaneous lesions suggestive of CL were sampled for parasitological diagnosis by direct examination (DE), culture in NNN medium, two kinetoplast DNA (kDNA) PCRs (Lmj4/Uni21 and 13A/13B primers) and one rRNA gene internal transcribed spacer 1 (ITS1) PCR (LITSR/L5.8S primers). The techniques were statistically analyzed and compared.Results: According to our consensus positive, 44 out of 58 samples were true positives. The 13A/13B-PCR and ITS1-PCR showed the highest sensitivities (100%). Parasite microscopy and culture detected 43% and 29% of the true positives, respectively, while culture and microscopy together improved sensitivity to 52%. PCRs 13A/13B and ITS1 were associated to four and one false positives, respectively, while the other assays were 100% specific. Furthermore, the ITS1-PCR-RFLP assay clearly identified the Leishmania species for all the true positives (44/44), whereas Lmj4/Uni21-PCR identified 35/44 samples. The comparison between the Leishmania molecular characterizations and the expected species according to the national data from the Ministry of Health indicate 7 discrepant results.Conclusions: The PCR-based assays tested on our samples increased the speed and sensitivity of the diagnosis of CL compared to the conventional techniques. Furthermore, we showed that we can not base the species identification on the national data from the Ministry of Health. Finally, we suggest the use of PCR-ITS1-RFLP for diagnosis and simultaneous identification of the species in the Moroccan epidemiological context, but also in similar areas of the Mediterranean Basin. © 2014 Mouttaki et al.


Drira I.,University of Sfax | Neji S.,Laboratory of parasitology mycology | Hadrich I.,University of Sfax | Sellami H.,University of Sfax | And 3 more authors.
Journal de Mycologie Medicale | Year: 2015

Trichophyton erinacei is a zoonotic fungus affecting hedgehogs. Although several human infections with this organism have been documented in the literature, it has rarely been isolated as a human pathogen. This paper reports on an erythematous lesion spotted on the hand of a 10-year-old girl. Based on the culture of the patient's skin scrapings, the pathogen was mycologically identified as T. erinacei, which was further confirmed by sequencing the internal transcribed spacers of the fungal nuclear ribosomal DNA using universal primer ITS1-ITS4. This is the first case of T. erinacei in a Tunisian patient. A survey was carried out on the environment of our patient, and the results revealed the presence of hedgehogs with suspect scaly lesions. The same fungus was isolated from the hair and scales of the hedgehog, which was confirmed by PCR sequencing. The frequency of T. erinacei has often been underestimated, which is attributed not only to the gaps of knowledge still existing in the current understanding of the dermatophyte but also to differential diagnosis problems. Molecular study offers a simple and rapid tool to identify the source of infection and, hence, avoid the risk of recurrence. © 2015 Elsevier Masson SAS.


PubMed | University of Sfax and Laboratory of parasitology mycology
Type: Case Reports | Journal: Journal de mycologie medicale | Year: 2015

Trichophyton erinacei is a zoonotic fungus affecting hedgehogs. Although several human infections with this organism have been documented in the literature, it has rarely been isolated as a human pathogen. This paper reports on an erythematous lesion spotted on the hand of a 10-year-old girl. Based on the culture of the patients skin scrapings, the pathogen was mycologically identified as T.erinacei, which was further confirmed by sequencing the internal transcribed spacers of the fungal nuclear ribosomal DNA using universal primer ITS1-ITS4. This is the first case of T.erinacei in a Tunisian patient. A survey was carried out on the environment of our patient, and the results revealed the presence of hedgehogs with suspect scaly lesions. The same fungus was isolated from the hair and scales of the hedgehog, which was confirmed by PCR sequencing. The frequency of T.erinacei has often been underestimated, which is attributed not only to the gaps of knowledge still existing in the current understanding of the dermatophyte but also to differential diagnosis problems. Molecular study offers a simple and rapid tool to identify the source of infection and, hence, avoid the risk of recurrence.


Khadraoui N.,Laboratory of Parasitology Mycology | Kallel K.,Laboratory of Parasitology Mycology | Bouchami O.,National Bone Marrow Transplant Center | Bouchakoua M.,Laboratory of Parasitology Mycology | And 5 more authors.
Annales de Biologie Clinique | Year: 2011

Candida albicans is the most important cause of fungal infections in intensive care units. The aim of this work was to compare the profiles of C. albicans in order to specify their genetic polymorphism and to determine the origin of these infections. Thirty-five C. albicans strains were collected from different clinical samples of 12 patients and three health-workers in an intensive care unit (ICU) in Rabta hospital of Tunisia, between August 2007 and April 2008. After digestion with BssHII, the isolates were typed by pulsed field gel electrophoresis (PFGE). The PFGE profiles were analyzed using a visual method, which showed three PFGE types (A, B and C) and the dendrogram generated three clusters (clusters I to III). An average similarity coefficient of 0.83, suggests that isolates are related. Copyright 2010 American Association for Clinical Chemistry, Inc.


Khadraoui N.,Laboratory of Parasitology Mycology | Kallel K.,Laboratory of Parasitology Mycology | Bouchami O.,Intensive Care Unit | Bouchakoua M.,Laboratory of Parasitology Mycology | And 6 more authors.
Journal de Mycologie Medicale | Year: 2010

Objective: The aim of our study was to compare some Candida tropicalis clinical isolates in order to specify their genetic polymorphism. Patients: The isolates were collected between August 2007 and April 2008 from 15 patients hospitalized in an intensive care unit in Rabta hospital, Tunisia. Materials and methods: All C. tropicalis isolates collected from patients hospitalized in this hospital unit were typed using pulsed field gel electrophoresis (PFGE). Results: During the study period, 15 patients revealed to be positive for C. tropicalis and 34 isolates were collected. Among them, 29 isolates belonged to the same PFGE type designated A (with three subtypes A1, A2 and A3), while two isolates belonged to the PFGE type B and the remaining three strains to the PFGE type C. Conclusion: The majority of isolates belonged to the same PFGE type (85%) but the subtype A1 was more represented than the subtypes A2 and A3. © 2010 Elsevier Masson SAS.

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