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1.
Med Mycol ; 53(5): 462-9, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25841053

RESUMO

Utilizing matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectra for Candida glabrata typing would be a cost-effective and easy-to-use alternative to classical DNA-based typing methods. This study aimed to use MALDI-TOF for the typing of C. glabrata clinical isolates from various geographical origins and test its capacity to differentiate between fluconazole-sensitive and -resistant strains.Both microsatellite length polymorphism (MLP) and MALDI-TOF mass spectra of 58 C. glabrata isolates originating from Marseilles (France) and Tunis (Tunisia) as well as collection strains from diverse geographic origins were analyzed. The same analysis was conducted on a subset of C. glabrata isolates that were either susceptible (MIC ≤ 8 mg/l) or resistant (MIC ≥ 64 mg/l) to fluconazole.According to the seminal results, both MALDI-TOF and MLP classifications could highlight C. glabrata population structures associated with either geographical dispersal barriers (p < 10(-5)) or the selection of antifungal drug resistance traits (<10(-5)).In conclusion, MALDI-TOF geographical clustering was congruent with MPL genotyping and highlighted a significant population genetic structure according to fluconazole susceptibility in C. glabrata. Furthermore, although MALDI-TOF and MLP resulted in distinct classifications, MALDI-TOF also classified the isolates with respect to their fluconazole susceptibility profile. Further prospective studies are required to evaluate the capacity of MALDI-TOF typing to investigate C. glabrata infection outbreaks and predict the antifungal susceptibility profile of clinical laboratory isolates.


Assuntos
Antifúngicos/farmacologia , Candida glabrata/isolamento & purificação , Candidíase/epidemiologia , Farmacorresistência Fúngica , Fluconazol/farmacologia , Técnicas de Tipagem Micológica , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Candida glabrata/química , Candida glabrata/classificação , Candida glabrata/efeitos dos fármacos , Candidíase/microbiologia , Análise por Conglomerados , França/epidemiologia , Genótipo , Humanos , Testes de Sensibilidade Microbiana , Repetições de Microssatélites , Tipagem Molecular , Fenótipo , Topografia Médica , Tunísia/epidemiologia
2.
J Mass Spectrom ; 50(2): 371-7, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25800019

RESUMO

Candida albicans is the most frequent yeast involved in human infections. Its population structure can be divided into several genetic clades, some of which have been associated with antifungal susceptibility. Therefore, detecting and monitoring fungal clones in a routine laboratory setting would be a major epidemiological advance. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectra results are now widely used as bar codes to identify microorganisms in clinical microbiology laboratories. This study aimed at testing MALDI-TOF mass spectra bar codes to identify clades among a set of C. albicans isolates. Accordingly, 102 clinical strains were genotyped using 10 microsatellite markers and analyzed via MALDI-TOF mass spectrometry. The mass spectra were compared with a reference spectral library including 33 well-characterized collection strains, using a Microflex(TM) system and Biotyper(TM) software, to test the capacity of the spectrum of a given isolate to match with the reference mass spectrum of an isolate from the same genetic clade. Despite high confidence species identification, the spectra failed to significantly match with the corresponding clade (p = 0.74). This was confirmed with the MALDI-TOF spectra similarity dendrogram, in which the strains were dispersed irrespective of their genetic clade. Various attempts to improve intra-clade spectra recognition were unsuccessful. In conclusion, MALDI-TOF mass spectra bar code analysis failed to reliably recognize genetically related C. albicans isolates. Further studies are warranted to develop alternative MALDI-TOF mass spectra analytical approaches to identify and monitor C. albicans clades in the routine clinical laboratory.


Assuntos
Candida albicans/genética , DNA Fúngico/análise , Repetições de Microssatélites/genética , Tipagem Molecular/métodos , Micologia/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Candida albicans/classificação , Candidíase/microbiologia , DNA Fúngico/química , DNA Fúngico/genética , Humanos
3.
Tunis Med ; 93(8-9): 543-7, 2015.
Artigo em Francês | MEDLINE | ID: mdl-26815521

RESUMO

BACKGROUND: Malaria has been eliminated in Tunisia since 1979, but the country remains, like all other countries harboring the vector, exposed to the potential risk of resurgence. OBJECTIVES: Describe the clinical and epidemiological investigation of 4 cases of autochthonous malaria in July 2013 and report the main actions of regional and national response. METHODS: Retrospective descriptive survey of the 4 clinical observations as well as the study of the regional report data of basic health care for the region of Tunis in 2013. RESULTS: Febrile table concomitant for 4 Tunisian male patients, aged from 21 to 27 years old ; fortuitous discovery of Plasmodium falciparum when checking thrombocytopenia of patient 1 ; diagnosis in cascade of other cases following the epidemiological investigation and field consultation with clinicians ; 3 simple forms and a neuromalaria of favorable evolution ; negative entomological survey for anopheles ; elimination of imported malaria and blood-borne ; airport malaria highly probable. The response included the establishment of a regional and national monitoring unit, an information program aimed at health professionals concerned and public opinion, the involvement of health and entomology teams for the detection and census of potential larval habitats and the implementation of local mosquito eradication measures. CONCLUSION: The clinical vigilance and competent biologist's eye is necessary to prevent the resurgence of this disease. The epidemiological surveillance system should be maintained and kept as well as the food safety standards monitoring at the borders.


Assuntos
Doenças Endêmicas , Malária Falciparum/epidemiologia , Adulto , Humanos , Masculino , Estudos Retrospectivos , Tunísia/epidemiologia , Adulto Jovem
4.
Parasitol Res ; 113(7): 2435-43, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24825311

RESUMO

The results of entomological studies carried out in the governate of Monastir (Tunisia) in 2009-2010 (captures and emergences from muds) focusing on Culicoides species are presented in the present study. Identification of Culicoides at the species level is based on morphological characters, and a molecular study has been carried out based on mitochondrial DNA cytochrome C oxidase I gene (COI) and D1 and D2 domains of the 28S rDNA. The DNA sequences reported here are related to 10 species (on 25 known) of Culicoides described in Tunisia: Culicoides cataneii-gejgelensis, Culicoides circumscriptus, Culicoides imicola, Culicoides jumineri, Culicoides kingi, Culicoides langeroni, Culicoides newsteadi, Culicoides paolae, Culicoides puncticollis and Culicoides sahariensis. DNA sequencing of the COI gene and D1D2 domains discriminated all morphologically determined species. The choice of D1D2 domains considered as a conserved region is informative for Culicoides species identification. The molecular analyses of COI has grouped both C. circumscriptus, C. puncticollis within two clusters and C. newsteadi within five subclusters. However, C. newsteadi shows relatively deep intraspecific divergence using COI sequences.


Assuntos
Ceratopogonidae/classificação , DNA Mitocondrial/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , RNA Ribossômico 28S/genética , Animais , Ceratopogonidae/genética , Filogenia , Filogeografia , RNA Ribossômico 28S/efeitos dos fármacos , Análise de Sequência de DNA , Tunísia
5.
Acta Trop ; 124(3): 221-8, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22940099

RESUMO

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.


Assuntos
Doenças do Cão/epidemiologia , Isoenzimas/análise , Leishmania infantum/enzimologia , Leishmania major/enzimologia , Leishmaniose/epidemiologia , Leishmaniose/veterinária , Topografia Médica , Animais , Doenças do Cão/parasitologia , Cães , Humanos , Leishmania infantum/classificação , Leishmania infantum/isolamento & purificação , Leishmania major/classificação , Leishmania major/isolamento & purificação , Leishmaniose/parasitologia , Epidemiologia Molecular , Tunísia/epidemiologia
6.
Pathol Biol (Paris) ; 59(6): e119-23, 2011 Dec.
Artigo em Francês | MEDLINE | ID: mdl-19896289

RESUMO

The cutaneous leishmaniasis (CL) is a parasitic disease which represents a serious problem for the public health not only in Tunisia but also all over the world. Its diagnosis is based on the techniques which are usually used, direct examination and in vitro culture. Because of several factors, these techniques lack sensitivity. The molecular biology, which is indeed more rapid and more sensitive, has proved its effectiveness in diagnosis of the CL. There are two main aims for our research work. First, to show the contribution of the Polymerase Chain Reaction (PCR) during the diagnosis of CL (of course by comparing the results obtained when using this technique with those found through the direct examination); second, to compare the two pairs of primers which amplify the leishmanien gene coding for the 18s ribosomal sub-unit: the pair R221/R332 (PCR1) and the pair Lei70L/Lei70R (PCR2). Our work was carried out upon 299 samples. One hundred and eighty-eight of them were positive using the direct examination and/or the PCR and 111 were negative. Only two samples were positive using of course the direct examination in comparison with 74 which were positive when using only the PCR (PCR1 and/or PCR2). Among these 74 samples, 64 where positive using only PCR2 in comparison with two samples which were positive using only PCR1. The eight remaining samples were at once positive for the PCR1 and the PCR2. The PCR (notably the PCR2) has proved a more significant percentage of positivity in comparison with direct examination: 98.98% for the PCR and 60.6% for direct examination.


Assuntos
Primers do DNA , Leishmaniose Cutânea/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Comportamento de Escolha , Primers do DNA/química , Primers do DNA/farmacologia , DNA de Protozoário/análise , DNA de Protozoário/genética , DNA de Protozoário/isolamento & purificação , Feminino , Humanos , Lactente , Leishmaniose Cutânea/parasitologia , Masculino , Pessoa de Meia-Idade , Técnicas de Diagnóstico Molecular/normas , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase/normas , Sensibilidade e Especificidade , Tunísia , Adulto Jovem
7.
Pathol Biol (Paris) ; 59(4): 234-9, 2011 Aug.
Artigo em Francês | MEDLINE | ID: mdl-19481368

RESUMO

INTRODUCTION: Intestinal microsporidiosis is recognised as an important cause of opportunistic parasitosis in immunocompromised patients, especially HIV-infected patients. Enterocytozoon bieneusi is the common causal agent. The diagnosis of intestinal microsporidiosis has usually based on microscopic detection of the spores of microsporidia species in stool samples, requires additional staining techniques as Modified Weber's trichrome stain. However, the detection of the spores can be difficult and species determination, which is important for defining the appropriate treatment, is impossible. Polymerase chain reaction (PCR)-based methods have been successfully used for detection of microsporidian infections. They are more sensitive and are able to identify microsporidia species. The purpose of this study is to identify E. bieneusi to adapt treatment and assess the true prevalence of the intestinal microsporidiosis due to this species in compromised patients in Tunisia. PATIENTS AND METHODS: One hundred and eighteen stools from immunocompromised patients, with a symptomatology in favour of the intestinal microsporidiosis, were analysed using light microscopy after staining with Modified Weber's trichrome stain and PCR. RESULTS: Only four were positive by Modified Weber's trichrome stain whereas eleven stools were positive by PCR, giving a prevalence of 20% in HIV-infected patients and 5,35% in human immunodeficiency virus-negative patients. CONCLUSION: This study confirms the usefulness of PCR in the diagnosis of the intestinal microsporidiosis due to E. bieneusi. Indeed, PCR has greater sensitivity than Modified Weber's trichrome stain and can identify the species of microsporidia in order to adapt the treatment.


Assuntos
Enterocytozoon/genética , Enterocytozoon/isolamento & purificação , Fezes/microbiologia , Hospedeiro Imunocomprometido , Microsporidiose/microbiologia , Reação em Cadeia da Polimerase , Infecções Oportunistas Relacionadas com a AIDS/microbiologia , Adolescente , Adulto , Feminino , Infecções por HIV/complicações , Infecções por HIV/microbiologia , Humanos , Enteropatias/microbiologia , Masculino , Microsporidiose/complicações , Microsporidiose/epidemiologia , Tunísia/epidemiologia
8.
Med Trop (Mars) ; 70(3): 269-73, 2010 Jun.
Artigo em Francês | MEDLINE | ID: mdl-20734597

RESUMO

BACKGROUND: Mycetoma is a chronic infection of cutaneous and subcutaneous tissue that can be caused by fungi or bacteria. It is endemic in tropical and subtropical areas but rare in Tunisia. PURPOSE: The purpose of this report is to describe epidemioclinical features, treatment and outcomes in patients presenting mycetoma in Tunisia. PATIENTS AND METHODS: The files of all patients treated for mycetoma in the Dermatology Department of La Rabta Hospital in Tunisia from 1982 to 2006 were retrospectively reviewed. RESULTS: A total of 15 cases of mycetoma were recorded during the study period. There were 6 men and 9 women with a mean age of 53.2 years. The most common clinical presentation was infiltrated erythematous plaques with sinus tracts (fistulas). Lesions were located on the foot in 12 cases. Thirteen patients reported the presence of grains in fluid discharging from fistulas. The cause of mycetoma was actinomycetes, i.e., Actinomadura madurae, in 9 cases and fungus in 6 cases including 3 due to Madurella mycetomi and 2 to Pseudallesheria boydii. Treatment was based on oral antibiotics for actinomycetoma and oral antifungals for eumycetoma. CONCLUSIONS: Mycetoma in Tunisia is still uncommon with a slight female predominance. The foot is the most frequent location. Diagnosis can be accomplished by direct mycologic examination, culture, and histololgy. There is no consensus on treatment that is often prolonged with numerous relapses.


Assuntos
Actinobacteria/isolamento & purificação , Micetoma/diagnóstico , Micetoma/epidemiologia , Adulto , Idoso , Antibacterianos/uso terapêutico , Antifúngicos/uso terapêutico , Quimioterapia Combinada , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Micetoma/tratamento farmacológico , Micetoma/microbiologia , Prevalência , Estudos Retrospectivos , Fatores de Risco , Dermatopatias Bacterianas/diagnóstico , Dermatopatias Bacterianas/epidemiologia , Resultado do Tratamento , Tunísia/epidemiologia
9.
Med Trop (Mars) ; 70(2): 135-6, 2010 Apr.
Artigo em Francês | MEDLINE | ID: mdl-20486346

RESUMO

Human cutaneous myiasis is a common dermatosis in tropical zones. The purpose of this report is to describe the first imported case of furuncular myiasis caused by Dermatobia hominis (human botfly) in Tunisia. The patient was a man returning from Bolivia. Furuncular myiasis was suspected based on epidemiological data and clinical examination showing pruriginous elevated lesions. Diagnosis was confirmed by identification of Dermatobia hominis larvae. Treatment was based mainly on manual removal of larvae. Since furuncular myiasis is unknown in Tunisia, it is important to remember this parasitic disease in differential diagnosis in patients presenting boil-like inflammatory papules following travel to Latin America.


Assuntos
Dípteros/patogenicidade , Miíase/diagnóstico , Adulto , Animais , Bolívia , Diagnóstico Diferencial , Dípteros/fisiologia , Feminino , Humanos , América Latina , Masculino , Oviposição , Viagem , Tunísia
10.
Sante ; 20(1): 21-9, 2010.
Artigo em Francês | MEDLINE | ID: mdl-20483704

RESUMO

INTRODUCTION: Intestinal microsporidiosis is an opportunistic parasitological infection affecting mainly immunocompromised patients, particularly those infected with HIV. PURPOSE: The purpose of this study was to analyse the epidemiological and clinical characteristics of intestinal microsporidiosis and the treatments available for it. MATERIAL AND METHODS: This retrospective study examined records collected over a 13-year period (from January 1995 through December 2007). It included 572 immunocompromised patients (279 HIV-infected patients and 293 without HIV infection) with symptoms suggesting intestinal microsporidiosis. All were tested systematically for microsporidia spores by modified (Weber's) Trichrome staining. RESULTS: Fourteen patients (10 men, 4 women) were diagnosed with intestinal microsporidiosis, for a prevalence of 2.4% overall, 3.6% in HIV-infected patients and 1.4% in those without HIV infection. Intestinal microsporidiosis affected 10 HIV-infected patients, 70% of whom had a CD4 count <100 cells/mm3. Their mean age was 30+/-15 years (range: 15 months to 48 years). The average age of HIV-infected patients (36 years) was significantly higher than of those without HIV infection (15 years). Thirteen patients had symptoms, most frequently diarrhea (11 cases), sometimes associated with dehydration (5 cases). Eight patients (57%) received only symptomatic treatment, and 4 (28.6%) received albendazole. No treatment was recommended in 2 cases (14.3%). Clinical course was marked by improvement in 6 cases, death in 5, and persistence of asymptomatic carriage in one. Two patients were lost to follow-up. CONCLUSION: Intestinal microsporidiosis is a parasitological disease that mainly affects AIDS patients with CD4 counts <100 cells/mm3. Its diagnosis requires special techniques. Its symptomatology is dominated by chronic diarrhea that can cause dehydration. Effective treatment requires identification of the species.


Assuntos
Enteropatias/epidemiologia , Microsporidiose/epidemiologia , Adolescente , Adulto , Criança , Feminino , Infecções por HIV/complicações , Infecções por HIV/microbiologia , Humanos , Hospedeiro Imunocomprometido , Incidência , Enteropatias/microbiologia , Masculino , Microsporum , Pessoa de Meia-Idade , Prevalência , Estudos Retrospectivos , Fatores de Risco , Estações do Ano , Tunísia/epidemiologia
11.
Ann Biol Clin (Paris) ; 67(3): 325-32, 2009.
Artigo em Francês | MEDLINE | ID: mdl-19411235

RESUMO

This is a retrospective study including 17 patients with rhino-orbito-cerebral mucormycosis diagnosed in a period of 16 years, between 1992 and 2007, in 8 men and 9 women. All patients were diabetic with ketoacidosis diabetes in 8 cases. Necrosis facial and ophthalmic symptoms were the most frequent presenting manifestations. The diagnosis was confirmed by mycological examination, with or without histopathology, identifying Rhizopus oryzae in 12 cases. Treatment consisted in systemic amphotericin B combined with surgical treatment in only 7 cases. The mortality rate was high (65%) due principally to the delay in diagnosis and absence of surgical treatment.


Assuntos
Anfotericina B/uso terapêutico , Complicações do Diabetes/microbiologia , Mucormicose/complicações , Antifúngicos/uso terapêutico , Complicações do Diabetes/tratamento farmacológico , Complicações do Diabetes/patologia , Complicações do Diabetes/cirurgia , Cetoacidose Diabética/tratamento farmacológico , Cetoacidose Diabética/patologia , Feminino , Humanos , Masculino , Mucormicose/tratamento farmacológico , Mucormicose/patologia , Mucormicose/cirurgia , Doenças Nasais/tratamento farmacológico , Doenças Nasais/microbiologia , Doenças Nasais/patologia , Doenças Nasais/cirurgia , Doenças Orbitárias/tratamento farmacológico , Doenças Orbitárias/microbiologia , Doenças Orbitárias/patologia , Doenças Orbitárias/cirurgia , Doenças dos Seios Paranasais/tratamento farmacológico , Doenças dos Seios Paranasais/microbiologia , Doenças dos Seios Paranasais/patologia , Doenças dos Seios Paranasais/cirurgia , Estudos Retrospectivos , Rhizopus/isolamento & purificação , Tunísia/epidemiologia
12.
Med Mal Infect ; 39(12): 914-9, 2009 Dec.
Artigo em Francês | MEDLINE | ID: mdl-19359114

RESUMO

Neuromeningeal cryptococcosis is a serious infection witch occurs essentially in immunodepressed patients and especially AIDS patients. We report 22 cases of cryptococcosis meningitis confirmed by the parasitology laboratory, in the Tunis Rabta hospital, over a 16-year period. Sixteen patients were HIV infected and six were not HIV infected. The clinical examination documented fever and headache as well as focal neurological signs especially in HIV infected patients. The mycological examination of CSF proved the diagnosis of neuromeningeal cryptococcosis in all cases. The first line treatment was Amphotericin B in 13 cases, Amphotericin B and 5Fluorocytosine in three cases, and fluconazole in six cases. 14 patients died, seven recovered, and one was lost to follow-up.


Assuntos
Meningite Criptocócica/epidemiologia , Infecções Oportunistas Relacionadas com a AIDS/tratamento farmacológico , Infecções Oportunistas Relacionadas com a AIDS/epidemiologia , Adulto , Anfotericina B/uso terapêutico , Antifúngicos/uso terapêutico , Comorbidade , Feminino , Fluconazol/uso terapêutico , Humanos , Hospedeiro Imunocomprometido , Masculino , Meningite Criptocócica/tratamento farmacológico , Pessoa de Meia-Idade , Tunísia/epidemiologia , Adulto Jovem
13.
Pathol Biol (Paris) ; 57(5): 373-7, 2009 Jul.
Artigo em Francês | MEDLINE | ID: mdl-19038508

RESUMO

Diagnosis of pneumocystis pneumonia is usually based on clinical features and X-rays photography and confirmed in the laboratory by visualisation of Pneumocystis organisms in stained preparations of respiratory specimens using several techniques (Gomori-Grocott, May-Grünwald Giemsa, bleu de toluidine O). Actually, PCR has considerably increased sensitivity of detection of Pneumocystis. The aim of this study is to compare conventional PCR results to those of staining techniques (Gomori-Grocott, May-Grünwald Giemsa) in addition to the X-ray and clinical findings in order to evaluate the contribution of each method. Sixty-four respiratory specimens were collected from 54 immuno-compromised patients with clinical symptoms of pulmonary infection. We diagnosed pneumocystis pneumonia in 16 patients according to staining techniques and/or typical clinical and radiological findings and/or response to treatment. Of the 15 patients, 14 were positive by PCR and only five were positive by direct examination, yielding a sensitivity and specificity of 93.3 and 87.1% for PCR and 33.3 and 100% for staining techniques. Conventional PCR provides a sensitive and objective method for the detection Pneumocystis jiroveci from less invasive sample.


Assuntos
Pneumocystis carinii/isolamento & purificação , Pneumonia por Pneumocystis/diagnóstico , Reação em Cadeia da Polimerase , Coloração e Rotulagem/métodos , Infecções Oportunistas Relacionadas com a AIDS/diagnóstico , Infecções Oportunistas Relacionadas com a AIDS/diagnóstico por imagem , Infecções Oportunistas Relacionadas com a AIDS/microbiologia , Adolescente , Adulto , Líquido da Lavagem Broncoalveolar/microbiologia , Corantes , DNA Fúngico/análise , Amarelo de Eosina-(YS) , Reações Falso-Positivas , Feminino , Neoplasias Hematológicas/complicações , Humanos , Hospedeiro Imunocomprometido , Síndromes de Imunodeficiência/complicações , Lactente , Masculino , Metenamina , Azul de Metileno , Pessoa de Meia-Idade , Pneumonia por Pneumocystis/diagnóstico por imagem , Pneumonia por Pneumocystis/microbiologia , Radiografia , Cloreto de Tolônio , Adulto Jovem
14.
Parasite ; 15(2): 143-50, 2008 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18642507

RESUMO

There has been a steady increase of visceral leishmaniasis during the past 20 years in Tunisia. In this study, we assess the value of two optimised PCR versus those of classical methods for the diagnosis of human visceral leishmaniasis. 106 samples were collected from 53 cases of pediatric visceral leishmaniasis. Peripheral blood and bone marrow samples were analysed both by parasitological methods (direct examination, leukocytoconcentration (LCC) and culture) and by PCR methods with two primer pair (R221/R332 and Lei 70L/Lei 70R). We diagnosed visceral leishmaniasis in all patients: 44 cases were diagnosed by culture (83%), 42 by direct examination of bone marrow (79%), 17 by LCC (32%), and 53 positive cases with both PCR assays (R221/R332 and/or Lei 70L/Lei 70R) (100 %). Regarding each PCR assay, for blood samples, the difference between the sensitivities of PCR Lei 70L/Lei 70R (86,8%) and PCR R221/R332 (17 %) is statistically significant with p-value 0.025. For bone marrow, the sensitivities of the two PCR methods were respectively 96,2% (Lei 70L/Lei 70R) and 75,5% (R221/R332). On the whole, PCR Lei 70L/Lei 70R was more effective than PCR R221/R332 and conventional methods for the two biological samples. Moreover, the requirement of less invasive sample using blood has the advantage of being repeatable for screening and for post therapeutic monitoring.


Assuntos
Medula Óssea/parasitologia , Leishmania/isolamento & purificação , Leishmaniose Visceral/diagnóstico , Reação em Cadeia da Polimerase/normas , Testes Sorológicos/normas , Animais , Criança , Humanos , Parasitemia/diagnóstico , Reação em Cadeia da Polimerase/métodos , Sensibilidade e Especificidade , Testes Sorológicos/métodos , Tunísia
15.
Bull Soc Pathol Exot ; 101(1): 29-31, 2008 Feb.
Artigo em Francês | MEDLINE | ID: mdl-18432004

RESUMO

Three clinico-epidemiological forms of cutaneous leishmaniasis (CL) were described in Tunisia: the zoonotic CL (ZCL) epidemic which occurred in the centre of the country caused by Leishmania major MON-25, the chronic CL (CCL) In the south-east of the country caused by Leishmania killicki MON-8 and the sporadic CL In the North (SCL) caused by Leishmania infantum MON-24. The latter form, described in 1991, prevails in northern Tunisia with approximately thirty cases per year. Its vector, unknown for a long time could be according to the last publications, Phlebotomus perfiliewi or Phlebotomus langeroni; however, its reservoir remains unknown until now. The systematic isoenzymatic characterization permits to identify a great number of strains improving then knowledge on the eco-epidemiology of the disease. Indeed, changes were noted in the geographical distribution of these clinical forms: extension of the ZCL to the North and South, extension of the CCL to North and the SCL to the centre. We report in this note the first mention of L. infantum MON-24 in the two provinces of the centre of Tunisia: Kairouan and Sidi Bouzid, confirming the extension of the SCL to the Centre.


Assuntos
Leishmania infantum/classificação , Leishmaniose Cutânea/parasitologia , Adolescente , Animais , Pré-Escolar , Dermatoses Faciais/parasitologia , Feminino , Humanos , Focalização Isoelétrica , Isoenzimas/análise , Leishmania infantum/isolamento & purificação , Úlcera Cutânea/parasitologia , Tunísia , Extremidade Superior/parasitologia
16.
Acta Trop ; 106(2): 132-6, 2008 May.
Artigo em Inglês | MEDLINE | ID: mdl-18402923

RESUMO

The different clinical forms of leishmaniasis are the result of both the immunological status of individuals and the species of the parasite causing the infection. In Mediterranean countries, the Leishmania infantum complex groups zymodemes which are responsible for visceral, cutaneous and exceptionally cutaneomucosal or mucosal leishmaniasis. We report in this study a synthesis concerning 254 cases of L. infantum that have been characterized at the "Laboratoire de Parasitologie" of the Rabta Hospital. The strains were isolated from human cases of visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL) by culture on NNN medium: 156 VL cases and 98 CL cases. The isoenzymatic characterization revealed three zymodemes of L. infantum. * L. infantum MON 1, a common zymodeme of VL,occurred in 154 cases (61%): 147 VL (95%) and 7 CL (5%). All CL cases were from the northern provinces, six of them occurring during an epidemic disease in 2001. * L. infantum MON 24, a common zymodeme of CL in the north, occurred in 98 cases (38.5%): 91 CL (93%) and 7 VL (7%). The seven VL cases were immunocompetent children aged from 8 months to 9 years and native of northern Tunisia. Two of the CL cases were from central regions of the country. This is the first time that cases from these regions are reported. * L. infantum MON 80, an uncommon zymodeme in Tunisia, occurred in two VL cases (0.5%): two children aged 7 and 5. The small number of strains of this zymodeme does not allow understanding of its epidemiological role. The results of this study indicate a low enzymatic variability of L. infantum in the country. However, our study includes only human strains and should be extended to animal ones (dogs, rodents and sand flies). This would lead to a better understanding of the epidemiology of leishmaniasis in Tunisia.


Assuntos
Isoenzimas/análise , Leishmania infantum/classificação , Leishmania infantum/enzimologia , Leishmaniose Cutânea/parasitologia , Leishmaniose Visceral/parasitologia , Adolescente , Adulto , Idoso , Animais , Criança , Pré-Escolar , Cães , Feminino , Humanos , Lactente , Focalização Isoelétrica/métodos , Leishmania infantum/isolamento & purificação , Leishmaniose Cutânea/epidemiologia , Leishmaniose Visceral/epidemiologia , Masculino , Pessoa de Meia-Idade , Proteínas de Protozoários/análise , Tunísia/epidemiologia
18.
Bull Soc Pathol Exot ; 100(4): 282-6, 2007 Oct.
Artigo em Francês | MEDLINE | ID: mdl-17982860

RESUMO

The purpose of this study was to evaluate in a retrospective analysis, cases of Mediterranean visceral leishmaniasis (VL) diagnosed in adults during a 20-year period in a department of infectious diseases. Demographic data, clinical and laboratory features and therapeutic findings were considered. During the study period, 22 cases of VL were diagnosed, and 6 (27%) were associated with HIV infection. Fever and splenomegaly were observed in all cases. Anaemia was constant. The anti-leishmanial IF titer was positive among 21 patients (95%). Smears from bone marrow aspiration were positive at microscopy in 95% of cases. Zymodeme analysis was carried out in nine isolates. L. infantum zymodeme MON-1 was characterized in all cases. Seventeen patients (77%) received meglumine antimoniate (MA) (20 mg SbV/kg per day) and 5 (23%) patients amphotericin B (AB) (0.5-1 mg/kg per day) for an average period of 25 days (10-49 days). Adverse events occurred in 7 patients (32%), among them 4 received AB. Clinical cure was achieved with success in 21 patients (95%). After a successful MA treatment of the initial episode, VL relapse was observed in one HIV-positive patient. Only one HIV-positive patient died from neurological disorders. VL is rare in adults. However, its incidence is increasing everywhere in the world, because of HIV-related cases. Its prognosis depends on the precocity of diagnosis and treatment.


Assuntos
Leishmaniose Visceral/epidemiologia , Adulto , Anfotericina B/uso terapêutico , Anemia/epidemiologia , Animais , Antimônio/uso terapêutico , Antiprotozoários/uso terapêutico , Febre/epidemiologia , Infecções por HIV/epidemiologia , Humanos , Leishmania infantum/classificação , Meglumina/uso terapêutico , Antimoniato de Meglumina , Compostos Organometálicos/uso terapêutico , Recidiva , Estudos Retrospectivos , Esplenomegalia/epidemiologia , Resultado do Tratamento , Tunísia/epidemiologia
19.
Ann Dermatol Venereol ; 134(10 Pt 1): 743-7, 2007 Oct.
Artigo em Francês | MEDLINE | ID: mdl-17978711

RESUMO

BACKGROUND: Onychomycosis is more frequent in the elderly. The aim of this Tunisian study was to determine the prevalence and the epidemiological, clinical and mycological characteristics of onychomycosis in elderly subjects. PATIENTS AND METHODS: This was a prospective study performed over a 7-month period (October 2005 - April 2006). The fingernails and toenails of 290 new consecutive patients aged 65 years and over presenting at general practitioners' clinics were examined by certified dermatologists. Clinical signs of onychomycosis were observed in 120 patients. 137 nail samples and 103 skin scrapings were obtained from these subjects. RESULTS: The study population comprised 51 men and 69 women (sex ratio M/F: 0.74) of mean age 72.33 +/- 6.2 years. Fingernails were involved in 28 cases (20.4%), toenails in 109 cases (79.5%), and both fingernails and toenails in 17 cases (12.4%). On average, patients had 2.7 infected fingernails and 3.9 infected toenails. The big toenail and thumbnail were affected most often respectively on the feet and on the hands. Among the clinical presentations seen, distolateral subungual onychomycosis was the most common clinical presentation, on both the feet and the hands. Mycologically confirmed onychomycosis was detected in 110 of 137 nail samples. The rate of positivity was 80.3%. The prevalence of onychomycosis was confirmed in 34.1% of patients examined (99/290). Dermatophytes were the main agents causing onychomycosis in our population (79%). The most frequently detected fungus was Trichophyton rubrum (72.1%). In toenail infections, dermatophytes were most frequently isolated (97%), while in fingernail infections, yeasts were most often isolated (75%), with Candida albicans being dominant (60%). No moulds were isolated in either the hands or the feet. The same fungus was isolated in both skin scrapings and nail samples from 33 patients. Trichophyton rubrum was detected in 28 cases. CONCLUSION: This study confirms a higher prevalence of onychomycosis in the elderly. It shows dermatophytes, in particular Trichophyton rubrum, to be the most frequently isolated causative agents in onychomycosis in the elderly, followed by yeasts, with no moulds being detected.


Assuntos
Onicomicose/epidemiologia , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Prevalência , Estudos Prospectivos
20.
Pathol Biol (Paris) ; 55(10): 521-4, 2007 Dec.
Artigo em Francês | MEDLINE | ID: mdl-17888586

RESUMO

The number of visceral leishmaniasis (VL) cases is in continuous growth in Mediterranean countries. In Tunisia, in addition to the traditional infantile form, more and more cases in immunocompetent or immunocompromised adults have been reported. However, co-infection VL-HIV remains rare in Tunisia and diagnosis of all the cases up till now has been done using traditional techniques (serology, direct examination and culture of bone marrow). However, the last years, several studies proved the greatest sensitivity of PCR in VL diagnosis. We carried out a systematic detection of Leishmania in peripheral blood for 25 HIV infected patients (10 were asymptomatic, 6 presented a fever and/or a paleness and/or an asthenia, and 9 had an opportunist infection other than VL). In all cases, the culture on Novy-Nicolle-McNeal (NNN) medium was negative by the end of the month. Serology carried out for 22 patients was negative in IFI in 17 cases, positive at the 1/20 for four others and positive at the 1/40 for one patient (confirmed by Western Blot technique). A PCR using the primers Lei70L-Lei70R, specific of the gene of Leishmania infantum, allowed the display of the specific band of 345 bp for 17 samples. The higher sensitivity of PCR compared to conventional methods is subject to the difficulty of result interpretation in PCR positive testing among patients not having any other marker of the disease which raises the question of significance for this asymptomatic bearing.


Assuntos
Infecções por HIV/complicações , Infecções por HIV/parasitologia , Leishmania infantum/isolamento & purificação , Adolescente , Adulto , Animais , Sangue/parasitologia , Criança , Feminino , Heterossexualidade , Humanos , Masculino , Valores de Referência , Tunísia
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