M Ben Said

Institut Pasteur de Tunis, Tunis-Ville, Tūnis, Tunisia

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Publications (75)65.42 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: In the present study, we applied the combination of one-dimensional gel electrophoresis, immunoblot and nano liquidchromatography-tandem mass spectrometry (nanoLC-MS/MS) to identify potential immunogenic proteins of Toxoplasma gondii tachyzoites that can be used for the development of reliable assays in the serodiagnosis of acquired toxoplasmosis in immunocompetent subjects. For this purpose, we developed an immunoblot using soluble and membrane extracts of GT1 Toxoplasma gondii tachyzoites and tested 194 positive and 100 negative sera obtained from pregnant women. Five bands of soluble antigens (98 kDa, 36 kDa, 33 kDa, 32 kDa and 21 kDa) and 4 bands of membrane antigens (41 kDa, 35 kDa, 32 kDa and 30 kDa) were selected as the most valuable in terms of sensitivity and specificity. Among these bands, only 2 bands of soluble antigen (33 kDa and 32 kDa) and 2 bands of membrane antigen (32 kDa and 30 kDa) showed a specificity ≥ 90%. After mass spectrometry and bioinformatics analysis, 7 proteins were identified as potential markers for serodiagnosis of toxoplasmosis. These proteins are: SRS34A, GRA7, GRA1, DG32, MIC5, ROP5 and Toxofilin. These proteins showed a 86% to 100% homology with proteins of both VEG and ME49 strains of T. gondii and a 58% to 87% homology with Hammondia hammondi; and can be considered as attractive candidates for the development of an immunochromatography test that can be used for the rapid diagnosis of toxoplasmosis and as a confirmatory test when routine techniques give equivocal results.
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    ABSTRACT: Hospital environment is considered the main source of invasive aspergillosis (IA) in leukemic patients. This study aimed to describe Aspergillus colonisation in leukemic patients and their hospital environment and to test whether Aspergillus environmental contamination was associated with IA. For a 2-year period including 14-month renovation work, 91 acute leukaemia inpatients at the hematology department of University hospital in Sousse (Tunisia) were prospectively included. The incidence of probable IA (EORTC/MSG criteria) was 9.9%. Fifty-six Aspergillus were isolated from 53 (6.5%) of 811 sputa collected from 35 (38.5%) patients. Aspergillus spp. were isolated in 59.7% of 494 air samples and in 52.8% of 1579 surface samples taken in the patients' room. Aspergillus section Nigri (72.7%) was the most frequent. Aspergillus contamination peaked in autumn and winter on surface and in summer and autumn in air samples and was higher (P = 0.03) during the renovation work period. Multivariate analysis showed that for each Aspergillus section Nigri CFU airborne contamination IA risk increased by 1.05 (P = 0.04). In Tunisia, Aspergillus section Nigri and Flavi, but not Fumigati, are chiefly involved in IA. Our findings support swift implementation of airborne fungal contamination control measures in areas where immunocompromised patient are hospitalised. © 2015 Blackwell Verlag GmbH.
    Mycoses 03/2015; 58(6). DOI:10.1111/myc.12320 · 1.81 Impact Factor
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    ABSTRACT: Les mucormycoses sont des infections graves dues à des champignons filamenteux de l’ordre des Mucorales. Elles surviennent le plus souvent chez des patients immunodéprimés. Nous rapportons cinq cas de mucormycose chez des patients hospitalisés dans le service de Maladies Infectieuses de Sousse - Tunisie, entre 2000 et 2013. Il s’agissait de 4 hommes et une femme, d’âge moyen 60 ans. Trois patients étaient diabétiques et un patient avait une leucémie aigue. Les localisations de la mucormycose étaient rhino-cérébrale, rhino-orbitaire, auriculaire, pulmonaire et cutanée. Les Mucorales isolés étaient Rhizopus arrhizus dans 3 cas et Lichtemia dans 2 cas. Tous les patients étaient traités par amphotéricine B et 2 patients ont eu, en plus, un débridement chirurgical. Deux patients sont décédés et 2 ont gardé une paralysie faciale périphérique.
    12/2014; DOI:10.1016/j.nmni.2014.12.002
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    ABSTRACT: The overall performance of quantitative assays in the detection of anti-Toxoplasma IgG is satisfactory but discrepancies between assays are not uncommon especially when IgG concentrations are close to the limit of detection of the tests.The purpose of our study was to identify soluble and membrane antigens extracted from T. gondii tachyzoites by immunoblot in order to select the most relevant antigenic bands to be used for qualitative serodiagnosis of acquired toxoplasmosis.We selected 5 relevant bands (98, 36, 33, 32 and 21 kDa) with soluble antigens and 4 relevant bands (42, 35, 32, 30 kDa) with membrane antigens which gave high sensitivity and/or specificity in immunodiagnosis.The association on the same blot of at least 3 out of the 5 relevant bands in the soluble antigen immunoblot showed the highest sensitivity/specificity (97.4% / 99.0% respectively).Our results indicate that immunoblot using soluble tachyzoite extract with simultaneous detection of at least 3 out of the 5 bands (98, 36, 33, 32 and 21 kDa) represents a valuable test for serodiagnosis of acquired toxoplasmosis and should be further evaluated as a confirmatory test for sera which give discrepant results in quantitative assays.This article is protected by copyright. All rights reserved.
    Parasite Immunology 11/2014; 36(12). DOI:10.1111/pim.12139 · 1.85 Impact Factor
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    ABSTRACT: The determination of the accurate immune status of pregnant women is crucial in order to prevent congenital toxoplasmosis. Equivocal results with conventional serological techniques are not uncommon when IgG titers are close to the cut-off value of the test, so that a confirmatory technique is needed. For this purpose, we developed a homemade immunoblot (IB) using soluble extract of T. gondii tachyzoites and assessed it by testing 154 positive, 100 negative and 123 equivocal sera obtained from pregnant women. In order to select the more valuable bands in terms of sensitivity and specificity, we used the Youden Index (YI). The highest YIs were those given by the 32, 36, 98, 21 and 33 bands. The simultaneous presence on the same blot of at least 3 bands showed a much higher YI (0.964) and was adapted as a positivity criterion. The analysis of results showed that our homemade IB correlated well with the commercial LDBIO Toxo II IgG® kit recently recommended as a confirmatory test (96.74% of concordance).
    The Korean Journal of Parasitology 06/2014; Vol. 52(No. 4):1-7. DOI:10.3347/kjp.2014.52.4.1 · 0.97 Impact Factor
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    ABSTRACT: The determination of the accurate immune status of pregnant women is crucial in order to prevent congenital toxoplasmosis. Equivocal results with conventional serological techniques are not uncommon when IgG titers are close to the cut-off value of the test, so that a confirmatory technique is needed. For this purpose, we developed a homemade immunoblot (IB) using soluble extract of Toxoplasma gondii tachyzoites and assessed it by testing 154 positive, 100 negative, and 123 equivocal sera obtained from pregnant women. In order to select the more valuable bands in terms of sensitivity and specificity, we used the Youden Index (YI). The highest YIs were those given by the 32, 36, 98, 21, and 33 bands. The simultaneous presence on the same blot of at least 3 bands showed a much higher YI (0.964) and was adapted as the positivity criterion. The analysis of results showed that our homemade IB correlated well with the commercial LDBIO Toxo II IgG® kit recently recommended as a confirmatory test (96.7% of concordance).
    The Korean Journal of Parasitology 06/2014; 52(5):493-499.. DOI:10.3347/kjp.2014.52.5.493 · 0.97 Impact Factor
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    ABSTRACT: Although scarce, available data suggest that the epidemiology of invasive aspergillosis (IA) in North Africa differs from northern countries, where more than 80 % is caused by Aspergillus fumigatus. This study aimed at describing the epidemiology of IA in the region of Sousse, Tunisia, and at assessing the usefulness of the available diagnostic tools. For 2 years, clinical and mycological data were prospectively collected from 175 neutropenia episodes of 91 patients hospitalised in the haematology department at the Farhat Hached hospital in Sousse (Tunisia). Screening for galactomannan antigen was positive in 40 % of neutropenia episodes; Aspergillus PCR was positive in 42 % of the tested sera. Nine patients were classified as probable and two as possible IA according to the EORTC/MSG criteria. Twelve patients who prematurely died, had no CT scan and could not be classified. Fifty-six Aspergillus spp. were isolated in 53 (6.5 %) sputa collected from 35 (20 %) patients. The following species were identified with MALDI-TOF mass spectrometry and DNA sequencing: A. niger, 35 %; A. flavus, 38 %; A. tubingensis, 19 %; A. fumigatus, 4 %; A. westerdijkiae, 2 % and A. ochraceus, 2 %. Our findings highlight the epidemiological features of IA in Tunisia, which is characterised by the predominance of Aspergillus spp. from sections Nigri and Flavi.
    Mycopathologia 04/2014; DOI:10.1007/s11046-014-9742-8 · 1.55 Impact Factor
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    ABSTRACT: The resistance of Aspergillus species to antifungal is increasingly reported and the knowledge of the local epidemiology and antifungal susceptibility pattern is pivotal to define adequate treatment policies. Our study aimed to: 1) describe the in vitro antifungal susceptibility profile of the Aspergillus species isolated from patients with haematological malignancies in Tunisia; 2) compare the E-test and Sensititre Yeast-One assays for the detection of paradoxical growth and trailing effect, both phenotypes commonly exhibited by Aspergillus spp. upon exposure to caspofungin and 3) to evaluate the mortality rate in patients according to the causative Aspergillus species and the antifungal treatment. We tested amphotericin B, itraconazole, voriconazole, posaconazole and caspofungin against 48 Aspergillus isolates (17, A. niger; 18, A. flavus; 9, A. tubingensis; 1, A. westerdijkiae; and 1, A. ochraceus) with the E-test. Minimal inhibition concentrations were above the epidemiological cut-off values for amphotericin B in 67% of A. flavus strains; for caspofungin in 22% of A. flavus strains; and for itraconazole in 22% of A. tubingensis strains, voriconazole and posaconazole MICs were below the epidemiological cut-off values for all strains. When exposed to caspofungin, 42% of the strains exhibited trailing effect and 38% paradoxical growth. Trailing effect occurred in 61% of A. flavus strains and paradoxical growth in 62% of Aspergillus section Nigri strains. E-test and Sensititre Yeast-One assays were only fairly concordant for the detection of these phenotypes. Repeatability of both assays was high for trailing effect but poor for paradoxical growth. The relatively high frequency of amphotericin B resistant strains makes voriconazole best adapted as a first-line treatment of invasive aspergillosis from amphotericin B to voriconazole in this hospital.
    SpringerPlus 01/2014; 3(1):19. DOI:10.1186/2193-1801-3-19
  • Annales de Dermatologie et de Vénéréologie 12/2013; 140(12):S585. DOI:10.1016/j.annder.2013.09.503 · 0.67 Impact Factor
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    ABSTRACT: The performance values of available techniques used in serodiagnosis of toxoplasmosis are satisfactory but they raise problems of equivocal and discordant results for very low IgG titers. Recently marketed, LDBio-Toxo II IgG Western blot (IB) showed an excellent correlation with the dye test. We estimated the proportion of equivocal and discordant results between the enzyme immunoassay Platelia Toxo IgG (EIA-IgG) and fluorescent antibody test (FAT) and assessed the usefulness of the IB as a confirmatory test. Out of 2,136 sera collected from pregnant women, 1,644 (77.0%) tested unequivocally positive and 407 (19.0%) were negative in both EIA-IgG and FAT. The remaining 85 (4%) sera showed equivocal or discordant results. Among them, 73 (85.9%) were positive and 12 (14.1%) were negative in IB. Forty-one (89.1%) equivocal sera in EIA-IgG and 46 (86.8%) equivocal sera in FAT were positive in IB. Reducing the cut-off values of both screening techniques improved significantly their sensitivity in detecting very low IgG titers at the expense of their specificity. In conclusion, equivocal results in routine-used techniques and their discordance in determination of the immune status in pregnancy women were not uncommon. IB test appeard to be highly useful in these situations as a confirmatory technique.
    The Korean Journal of Parasitology 08/2013; 51(4):485-8. DOI:10.3347/kjp.2013.51.4.485 · 0.97 Impact Factor
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    I Dhib · A Fathallah · A Yaacoub · F Hadj Slama · M B Said · R Zemni
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    ABSTRACT: Onychomycosis is one of the most prevalent dermatophytic diseases. Mycological methods used in the conventional diagnosis may not be optimal. Multiplex (MX) PCR was reported as a reliable alternative. Dermatophyte gene sequence records were used to design a MX PCR for detection and identification of dermatophytes in nail specimens. A MX PCR method based on the amplification of the chitin synthase 1 and internal transcribed spacer genes was developed. The study included 93 strains of dermatophytes and non-dermatophytic fungi, six dermatophytic reference strains and 201 nail specimens from patients with dermatophytic onyxis. DNA extraction directly from nail samples was carried out by using the QIAamp DNA extraction kit (Quiagen). A set of primers was designed and their specificity was assessed. MX PCR detected the causal agent in specimens from which Trichophyton rubrum and T. interdigitale grew in culture and also identified a dermatophyte species in an additional 32 specimens that were negative in microscopy and culture. None of the investigated non-dermatophytic strains was positive. Sensitivity of MX PCR was higher as compared to mycological examination (97% vs. 81.1%). MX PCR for direct detection of dermatophytes from nail samples yielded mixed flora in 32.8% of samples. MX PCR proved sensitive and adequate for the diagnosis of dermatophytic onychomycosis. It is much adapted to cases where culture is negative or contaminated by overgrowing moulds, which makes the identification of the causal agent problematic.
    Mycoses 06/2013; 57(1). DOI:10.1111/myc.12096 · 1.81 Impact Factor
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    ABSTRACT: In Tunisia, despite the absence of a national program of congenital toxoplasmosis prevention, serodiagnosis is regularly carried out in pregnant women for prenatal screening. We report our experience on congenital toxoplasmosis over the last 11 years (2000–2011). Methods We reviewed the clinical, biological and therapeutic features of 21 cases of congenital toxoplasmosis. Results The infection occurred in the first trimester in four cases, in the second trimester in six cases and in the third trimester in four cases. Presumed date of maternal contamination could not be specified in seven cases. Spiramycin was prescribed in 16 cases and pyrimethamine-sulfadiazine in one case with a positive PCR. Fetal ultrasound was poorly contributive in the antenatal screening. It showed intra-uterine growth retardation in one case. At birth, 16 neonates were asymptomatic, four presented neurological symptoms and four presented ocular involvement. The postnatal serology was positive for 21 neonates during the first month of life. IgA were positive in 76.9 % of cases and IgM in 66.6 % of cases. Comparison of immunoblot profiles showed a neo-synthesis of IgG and/or IgM in 57.1 % of cases. Twenty neonates were treated with sulfadoxine-pyrimethamine. The evolution was marked by the early death of two neonates. Conclusion Severe congenital toxoplasmosis is still relevant in our country. The management of pregnant women at risk needs to be improved.
    Journal de Pédiatrie et de Puériculture 04/2013; 26(2):83–89. DOI:10.1016/j.jpp.2013.01.004
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    ABSTRACT: Purpureocillium lilacinum is a saprophytic fungus found in soil and decaying organic matter, but has been reported as an emerging pathogen in immunocompromised patients and following surgical procedures. Infections caused by this mold are often difficult to treat because of its intrinsic resistance to conventional antifungal agents and variable susceptibility to novel triazoles. In immunocompetent subjects, infections caused by P. lilacinum are unusual and mainly involve the skin. We describe herein a case of cutaneous hyalohyphomycosis due to this fungus in an immunocompetent girl without any predisposing risk factors and review the previously reported cases in immunocompetent hosts.
    Medical mycology: official publication of the International Society for Human and Animal Mycology 01/2013; 51(6). DOI:10.3109/13693786.2012.757656 · 2.26 Impact Factor
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    ABSTRACT: Candida albicans is one of the most medically important fungi because of its high frequency as a commensal and pathogenic microorganism causing superficial as well as invasive infections. Strain typing and delineation of the species are essential for understanding its biology, epidemiology and population structure. A wide range of molecular techniques have been used for this purpose including non DNA-based methods (multilocus enzyme electrophoresis), conventional DNA-based methods (electrophoretic karyotyping, random amplified polymorphic DNA, amplified fragment length polymorphism, restriction enzyme analysis with and without hybridization, rep-PCR) and DNA-based methods called exact typing methods because they generate unambiguous and highly reproducible typing data (including microsatellite length polymorphism and multilocus sequence typing). In this review, the main molecular methods used for C. albicans strain typing are summarized and their advantages and limitations are discussed with regard to their discriminatory power, reproducibility, cost and ease of performance. © 2013 The Authors Journal of Applied Microbiology © 2013 The Society for Applied Microbiology.
    Journal of Applied Microbiology 01/2013; 114(6). DOI:10.1111/jam.12132 · 2.39 Impact Factor
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    ABSTRACT: Geotrichum capitatum infection is uncommon, and has been exclusively reported in immunocompromised patients. The prognosis is poor with a mortality rate ranging from 50 to 90%. We report 3 cases of Geotrichum capitatum fungemia in neutropenic patients receiving chemotherapy for acute myeloblastic leukemia.The infection was successfully cured with voriconazole in 1 case and was fatal in the 2 remaining cases despite treatment with amphotericin B.
    Medical Mycology Case Reports 12/2012; 1(1):88–90. DOI:10.1016/j.mmcr.2012.09.003
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    ABSTRACT: Discrimination of the Old World Leishmania parasites is important for diagnosis and epidemiological studies of leishmaniasis. We have developed PCR assays that allow the discrimination between L. major, L. tropica and L. infantum Tunisian species. The identification was performed by a simple PCR targeting cysteine protease B (cpb) gene copies. These PCR can be a routine molecular biology tools for discrimination of Leishmania spp. from different geographical origins and different clinical forms. Our assays can be an informative source for cpb gene studying concerning drug, diagnostics and vaccine research. The PCR products of the cpb gene and the N-acetylglucosamine-1-phosphate transferase (nagt) Leishmania gene were sequenced and aligned. Phylogenetic trees of Leishmania based cpb and nagt sequences are close in topology, and present the classic distribution of Leishmania in the Old World. The phylogenetic analysis has enabled the characterization and identification of different strains, using both multicopy (cpb) and single copy (nagt) genes. Indeed, the cpb phylogenetic analysis allowed us to identify the Tunisian L. killicki species, and a group which gathers the least evolved isolates of the L. donovani complex, that was originated from East Africa. This clustering confirms the African origin for the visceralizing species of the L. donovani complex.
    Acta tropica 12/2012; 125(3). DOI:10.1016/j.actatropica.2012.11.012 · 2.52 Impact Factor
  • I Dhib · A Fathallah · A Yaacoub · R Zemni · R Gaha · M B Said
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    ABSTRACT: Onychomycosis is the most frequently encountered nail disease and may be difficult to diagnose and treat. The objective of this study was to determine the prevalence, the clinical and mycological characteristics of onychomycosis in central Tunisia. It is a retrospective study performed over a 22-year period (1986-2007). It included 7151 patients (4709 women and 2442 men) with suspected fingernails and/or toenails onychomycosis. The patients were referred to the Mycology-Parasitology Laboratory of Farhat Hached hospital in Sousse for mycological examination. Both direct microscopy and culture of the nail material were performed to diagnose and identify the causative fungal species. Onychomycosis was confirmed in 78.6% of investigated patients (5624/7151). The positivity rate was higher in women as compared with men. In both men and women, fingernails were most frequently involved than toenails. No significant relation was found between gender and toenails onychomycosis, whereas fingernails were frequently involved in women. As far as aetiological agents are considered, dermatophytes, yeast and moulds were responsible for 49.9%, 47.4% and 2.7% of onyxis cases respectively. In fingernail infections, yeast were the most frequent fungi (83.6%), Candida albicans being the leading species (51.6%). In contrast, in toenail infections, dermatophytes were more frequent (74.1%). Trichophyton rubrum was by far the dominant species (88.1%). Yeast were observed more frequently in women whereas dermatophytes were more common in men. Moulds were involved in 4.2% of cases. The most frequent species were Aspergillus sp. and Chrysosporium sp. Onychomycosis is a frequent disease in central Tunisia. T. rubrum is the predominant agent in toenails infection and yeast, mainly C. albicans, in fingernails onychomycosis.
    Mycoses 10/2012; 56(3). DOI:10.1111/myc.12016 · 1.81 Impact Factor
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    ABSTRACT: Objective The aim of our study was to evaluate the level of fungal contamination of food served to patients in the hematology unit of Farhat Hached hospital, Sousse (Tunisia).Methods We analyzed 159 food served to 90 patients during the post-chemotherapy aplasia period.ResultsThe overall rate of fungal contamination was 66.6%. Fruits and green salads’ contamination was very high (90 and 100%, respectively). Aspergillus species were the most frequent (48.4%) followed by Candida (25.1%), Penicillium (22%), Rhizopus (17.6%) and Cladosporium (17%). The presence of Aspergillus on fruits (66.6%) represents a real source of contamination by inhalation of spores and the presence of Candida (25.1%) in any type of food is a source of gastrointestinal colonization.Conclusion The diet of neutropenic patients should be submitted to a strict surveillance to reduce the risk of fungal infections of food.
    Nutrition Clinique et Métabolisme 09/2012; 26(3):114–118. DOI:10.1016/j.nupar.2012.05.001 · 0.62 Impact Factor
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    ABSTRACT: Onychomycosis is one of the most prevalent dermatophytic diseases. Mycological methods used in the conventional diagnosis may not be optimal. PCR was reported as a reliable alternative in the diagnosis of dermatophytosis. A PCR method based on the amplification of the chitin synthase 1 gene was developed. The study included 119 strains of dermatophytes and non dermatophytic fungi, eight dermatophytic reference strains and 201 nail specimens from patients with dermatophytic onyxis. DNA extraction was carried out by using the QIAamp DNA extraction kit (Quiagen). PCR positivity was based on the production of a specific 432bp fragment. None of the investigated non dermatophytic strains was positive. Sensitivity of PCR was higher as compared to mycological examination (90.5% vs. 81.1%). PCR was positive in 31 onyxis cases with positive direct examination but negative or contaminated culture. In contrast, PCR was negative in 10 cases where both direct examination and culture were found positive. PCR is an adequate tool for the diagnosis of dermatophytic onychomycosis. It is much adapted to cases where culture is negative or contaminated by overgrowing molds, which makes the identification of the causal agent problematic.
    Journal de Mycologie Médicale/Journal of Medical Mycology 09/2012; 22(3):249-55. DOI:10.1016/j.mycmed.2012.07.050 · 0.40 Impact Factor
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    ABSTRACT: Over a period of ten years, a series of 694 Leishmania strains from Tunisian leishmaniasis foci were isolated and identified by isoenzymatic analysis. Strains were obtained from human cutaneous and visceral leishmaniasis in immunocompetent subjects, visceral leishmaniasis in imunocompromised individuals and from dogs with visceral leishmaniasis. Two classically dermotropic species, Leishmania (L.) major and Leishmania killicki were found. L. major with the single zymodeme MON-25 was the most isolated in cutaneous leishmaniasis foci of the Centre and South of Tunisia with a recent northern extension. L. killicki zymodeme MON-8 was sporadically found both in its classical microfocus of Tataouine in southeastern Tunisia as well as in some new foci in Southwestern, Central and Northern Tunisia. Leishmania infantum with its three zymodemes MON-1, MON-24 and MON-80 was isolated from both visceral and cutaneous human cases. The majority of L. infantum strains were found in the Northern part of the country; however, some strains were reported for the first time in the Southern part. L. infantum MON-1 was the only zymodeme isolated from canine leishmaniasis.
    Acta tropica 08/2012; 124(3):221-8. DOI:10.1016/j.actatropica.2012.08.012 · 2.52 Impact Factor

Publication Stats

241 Citations
65.42 Total Impact Points

Institutions

  • 2014
    • Institut Pasteur de Tunis
      Tunis-Ville, Tūnis, Tunisia
  • 2009–2014
    • Centre Hôpital Universitaire Farhat Hached
      Susa, Sūsah, Tunisia
  • 2012
    • Ministère de la Santé Publique de Tunisie
      Tunis-Ville, Tūnis, Tunisia
  • 1989–1992
    • Faculty of Medecine of Tunis
      Tunis-Ville, Tūnis, Tunisia