The efficacy of voriconazole in 24 ocular Fusarium infections.

PURPOSE: We examined, retrospectively, the efficacy of voriconazole in Fusarium eye infections. METHODS: Voriconazole-treated patients with proven or probable keratitis or endophthalmitis from the voriconazole database (9 patients) and six French ophthalmology departments (15 patients) were included. Sociodemographic features, predisposing factors, history of corneal trauma, associated ocular conditions, other diseases and prior therapies were analysed. Investigator-determined success was defined as infection resolution with medical treatment. Failure was no response or persistent infection and required surgery. RESULTS: Most patients were Caucasian (83 %) and male (71 %). The infection was keratitis (63 %) or endophthalmitis (37 %) and proven in 23 (96 %). Prior therapy included topical and/or systemic amphotericin (46 %), fluconazole (17 %) or others (33 %), often in combination. Causative fungi were Fusarium solani (14, 58 %), Fusarium moniliforme (1), Fusarium oxysporum (1) and Fusarium spp. (8). Voriconazole was administered systemically, topically and/or by intraocular injection, and 16 patients (67 %) received salvage and eight primary therapy. The overall response was 67 % (73 % keratitis and 56 % endophthalmitis) but seven patients required adjunctive surgery. However, response was 63 % for eight primary therapy patients and 69 % for 16 salvage therapy patients. Response by species was Fusarium solani 64 % (9/14) and all others 80 % (8/10). In 13 patients (77 %), voriconazole was used in combination (response 69 vs. 64 % alone) with topical [amphotericin B 10/24 (42 %), caspofungin 5 (21 %), natamycin 1 (4 %)] and systemic agents [caspofungin 3 (13 %), amphotericin 2 (8 %)]. CONCLUSIONS: Topical and systemic voriconazole appears to be effective alone or in combination with other agents for treating severe Fusarium keratitis or endophthalmitis.

Human epithelial cell cultures from superficial limbal explants.

PURPOSE: To study the kinetics of growth and the phenotype of cells cultured from human limbal explants in a cholera toxin-free medium with no feeder cell layer. METHODS: Human organ-cultured corneas were used to prepare limbal explants (full-thickness and superficial limbal explants) and corneal stromal explants. Cell growth kinetics and phenotypes were assessed by cultivating explants in cholera toxin-free Green medium. Epithelial and progenitor cell markers were assessed by immunocytochemistry, flow cytometry, and Reverse Transcription and Polymerase Chain Reaction (RT-PCR). RESULTS: The successful epithelial cell growth rates from full thickness limbal explant and superficial limbal explant tissues were 41 and 86%, respectively (p=0.0001). The mean cell area and the percentage of small cells in superficial and full-thickness explant cultures were, respectively, 317 µm(2) and 429 µm(2), and 8.9% and 1.7% (p<0.001). The percentage of positive cells in superficial and full-thickness limbal explant cultures as assessed by immunocytochemistry were the following: broad spectrum cytokeratins (cytokeratins 4, 5, 6, 8, 10, 13, and 18 [MNF116]), 82%/37% (p=0.01); cytokeratin 3 (CK3), 74%/25% (p=0.009); cytokeratin 19 (CK19), 46%/25% (p=0.19); vimentin, 56%/53% (p=0.48); delta N p63α, 54%/0% (p<0.001); and ABCG2, 5%/0% (p=0.1). Flow cytometry showed a higher percentage of small cells, a higher percentage of MNF116+ cells, and stronger expression of progenitor-associated markers in superficial than in full-thickness explant cultures. For superficial limbal explant cultures, analysis of the expression profiles for various mRNAs at the end of 21 days of culture showed high levels of expression of the mRNAs encoding CK3, vimentin, and CK19. The expression of mRNA of delta N p63α and ABCG2 was weaker. Cultures obtained from full-thickness limbal explants featured no expression of mRNA of CK19, delta N p63α, and ABCG2, whereas mRNAs encoding CK3 and vimentin were detected. Human corneal stromal explants cultured with the same medium featured late cell growth, large mean cell area (2,529 µm(2)), no expression of cytokeratins, delta N p63α, and ABCG2, and high expression of vimentin. CONCLUSIONS: Superficial limbal explants appear to be superior to full-thickness limbal explants for growing human limbal epithelial cells. Preparation of explants using surgical facilities (i.e., operating microscope and microsurgical blades) led to a dramatic increase in the percentage of successful cultures, higher epithelial cell growth, decreased fibroblast contamination, and better preservation of limbal epithelial progenitors.

[New ultra rapid test for detection of bacteria]. / Nouveau test ultrarapide pour la détection de bactéries.

INTRODUCTION: Bacteriological testing is aimed to reduce the risk of transmission of infections. However, the detection of Bacteria by culture requires from 18hours to 14 days and may produce erroneous results for fastidious species. The goal of this work was to design and validate a new tool for bacterial testing. METHODS: The test is based on the fast real-time PCR (frt PCR). The DNA extracted from samples containing internal controls are introduced into four tubes containing primers and probes for the frt PCR. The cycling program consists in 1×at 95°C for 10min and 45×(15s at 95°C, 8s) at 52°C and 10s at 72°C. RESULTS: The frt PCR detects 0,01 CFU/µl of Bacteria and identifies eight Genera without interferences from the environment or from fungi and with no need for melting curve analysis or additional sequencing. DISCUSSION: The frt PCR detects and quantifies Bacteria identifying and assessing the load of Staphylococci, Streptococci, Haemophilus, Pseudomonas, Enterobacteria, Acinetobacter, Propionibacteriacae and Corynebacteria. CONCLUSION: Cultures require at least 24hours but the new frt PCR reduces the time to 90minutes. Larger series of samples are necessary to confirm the usefulness of this new test for routine bacterial sterility controls.

Microbiological efficacy of 3-day treatment with azithromycin 1.5% eye-drops for purulent bacterial conjunctivitis.

PURPOSE: Antibacterial efficacy of topically applied azithromycin 1.5% was compared with tobramycin 0.3% in a multicenter, randomized, investigator-masked study for the treatment of purulent bacterial conjunctivitis. METHODS: A total of 1043 adults and children received either azithromycin twice daily for 3 days (n=524) or tobramycin every 2 hours while awake for 2 days, then four times daily for 5 days (n=519). Conjunctival swabbing was taken at days 0, 3, and 9, using alginate swabs resuspended in a dissolution-transport medium, providing rapid and reproducible results. Cagle's criteria were used to define the pathogenicity level for each isolated bacterium. RESULTS: In the per-protocol set, the rate of bacteriologic resolution was 85.2% for azithromycin versus 83.8% for tobramycin on day 3, and 92.8% for azithromycin versus 94.6% for tobramycin on day 9. Azithromycin was demonstrated to be noninferior to tobramycin according to the 10% noninferiority margin. Although some bacteria were categorized as resistant to tested antibiotics, eradication was observed (for azithromycin: Acinetobacter, Enterobacteriaceae, Pseudomonas), highlighting the specific pharmacokinetics/pharmacodynamics of the ocular route. CONCLUSIONS: In total, topical therapy with azithromycin 1.5% administered only twice daily for 3 days effectively eradicates most pathogenic bacteria associated with bacterial conjunctivitis. These microbiologic results are in accordance with the observed clinical outcome. This new anti-infective product has the advantage of a short treatment course which could lead to an improvement in patient compliance.

Quality assessment of conjunctival specimens for detection of Chlamydia trachomatis by PCR in children with active trachoma.

One component of control programmes to eliminate trachoma is the treatment of Chlamydia trachomatis infection. A diagnosis of trachoma is based on clinical grounds, but the signs of active trachoma do not always correlate with the presence of C. trachomatis. During a therapeutic trial, the level of C. trachomatis infection in children with active trachoma in Guinea and Pakistan was assessed using a qualitative commercially available PCR that targeted the C. trachomatis plasmid. The influence of the quality of specimens on the efficiency of the PCR was investigated using two quantitative real-time PCRs targeting the specific omp1 gene of C. trachomatis and human chromosomal DNA, respectively. C. trachomatis was detected in c. 23% of children (aged 1-10 years) who presented with clinically active trachoma. Controls showed that PCR-related problems did not influence this detection rate. For 14% of the positive samples, C. trachomatis was detected in only one eye, with a significantly lower mean load of bacteria. These results suggest that epidemiological and therapeutic surveys should be conducted by sampling and testing both eyes. Moreover, the high variability of the cell load observed in the conjunctival swabs suggests that the effectiveness of swabbing may be questionable.

Detection by broad-range real-time PCR assay of Chlamydia species infecting human and animals.

BACKGROUND: Tests available for molecular diagnosis of chlamydial infections detect Chlamydiatrachomatis, but do not find other Chlamydia species associated with genital, ophthalmic, cardiovascular, respiratory or neurological diseases. The routine detection of all Chlamydia species would improve the prognosis of infected people and guide therapeutic choices. AIM: To design and validate a sensitive, specific, reproducible, inexpensive and easy-to-perform assay to quantify most Chlamydia species. METHODS: Primers and probe were selected using the gene coding for the 16S rRNA. The detection limits were assessed for suspensions of Chlamydia trachomatis, Chlamydia psittaci and Chlamydia pneumoniae. The performance of this test was compared with that of two commercial kits (Amplicor-Roche and Artus) on 100 samples obtained from children with trachoma. RESULTS: The detection capacities for Chlamydia trachomatis of the broad-range real-time polymerase chain reaction (PCR) were similar or slightly better than those obtained with commercial kits (0.2 copies of DNA/microl). Only the broad-range PCR identified specimens containing Chlamydia psittaci and Chlamydia pneumoniae. The commercial kits and the broad-range assay detected Chlamydia species in 5% and in 11%, respectively, of samples from children with trachoma. CONCLUSIONS: This new real-time PCR offers a sensitive, reproducible assay that produces results in <3 h. With panels of quantified Chlamydia species, this real-time PCR can be run with all real-time PCR equipment. Larger trials are needed to confirm the utility of this test in diagnosis and for therapeutic follow-up.

Effects of topical anaesthetics and fluorescein on the real-time PCR used for the diagnosis of Herpesviruses and Acanthamoeba keratitis.

BACKGROUND: The early microbiological diagnosis of corneal infections may prevent the condition from worsening. AIM: To study the potential interferences of oxybuprocain and fluorescein solutions used by ophthalmologists on the performances of the real-time polymerase chain reaction (PCR) carried out as routine test for diagnosis of keratitis. METHODS: Quantified suspensions of Herpes simplex virus (HSV1), Varicella zoster virus (VZV), Cytomegalovirus (CMV) and Acanthamoeba with and without oxybuprocain or fluorescein added before DNA extraction were tested by real-time PCR. RESULTS: The capacities of the real-time PCR to detect HSV, VZV, CMV and Acanthamoeba were reduced by oxybuprocain and fluorescein. Both products diluted to 1/16 reduced the PCR detection capacities for more than 2 logs (DNA copies/sample). CONCLUSIONS: The simultaneous introduction of fluorescein or topical anaesthetics into the tubes containing the specimens to be tested by PCR may lead to false negative results. Because corneal specimens for microbiological diagnosis of keratitis are obtained after topical administration of anaesthetics and corneal staining with fluorescein, ophthalmologists should be aware to rinse the eye surface intensively with appropriate eye solutions to minimise the risks of misdiagnosis.

[Postoperative endophthalmitis: 2000-2002 results in the XV-XX National Ophthalmologic Hospital]. / Infections nosocomiales endoculaires au CHNO des XV-XX de 2000 à 2002.

PURPOSE: Prevention of nosocomial infection is a priority for the Infection Control Committee (ICC). Following their recommendations, the XV-XX National Ophthalmologic Hospital's ICC conducted a survey on cases of infection and we report these results for the 2000-2002 period. METHODS: During the 2000-2002 period, 21,384 programmed intraocular surgeries were done. Every day, a hygiene coordinator was informed of each bacterial or fungal laboratory test on intraocular samples and for all patients who were hospitalized for endophthalmitis. After data analysis with a referent ICC physician, resulting infection was declared postoperative nosocomial endophthalmitis. For these patients, prophylactic antibiotic use, the surgery report, and bacterial laboratory test results were reviewed. RESULTS: The overall 3-year incidence of suspected postoperative endophthalmitis after intraocular surgery was 2.0 per 1000 (42 cases). Confirmed microbiological growth was demonstrated in 19 cases (45%). For cataract surgery, the incidence was 1.0 per 1000 for acute-onset culture-proven postoperative endophthalmitis, 0.1 per 1000 for delayed culture-proven postoperative endophthalmitis, 2.1 per 1000 for acute-onset suspected postoperative endophthalmitis and 0.5 per 1000 for delayed suspected postoperative endophthalmitis. Gram-positive cocci were isolated in 77%. Twenty-one patients received systemic prophylactic antibiotics. Bacterial growth was positive in five of these 21 cases. For two cases, organisms were resistant to the prophylactic antibiotics used. CONCLUSION: Monitoring postoperative nosocomial infection is mandatory to detect incidence variation and evaluate infection control management. Prophylactic antibiotic efficacy remains to be evaluated.

Homozygosity mapping of spinocerebellar ataxia with cerebellar atrophy and peripheral neuropathy to 9q33-34, and with hearing impairment and optic atrophy to 6p21-23.

With the availability of a simple molecular test that distinguishes Friedreich ataxia, the most frequent form of inherited ataxia, from other recessive ataxias, it now becomes possible to unravel the genetic heterogeneity of the latter. We have now localised two genes causing autosomal recessive spinocerebellar ataxia in two consanguineous families. In the first family, the four affected Japanese sibs had spinocerebellar ataxia associated with elevated levels of serum creatine kinase, gamma-globulin, and alpha-foetoprotein. Homozygosity over a 20 cM region allowed to demonstrate linkage at 9q33.3-34.3 with a lod score of 3.0. Genotyping two unrelated Japanese patients from first degree consanguineous parents revealed that one was homozygous for the same region but did not share the biochemical features. In the second family, an Israeli uncle and a niece were affected by an early-onset recessive ataxia and subsequently developed hearing impairment and optic atrophy. Homozygosity over a 17 cM region allowed demonstration of linkage at 6p21-23 with a lod score of 3.25. These two localisations of autosomal recessive ataxia genes represent a first step toward the identification of genetically homogenous, non-Friedreich, ataxic patients and subsequent cloning of the genes.

Genomic organization of the MTM1 gene implicated in X-linked myotubular myopathy.

X-linked recessive myotubular myopathy (XLMTM) is a very severe congenital muscular disease characterised by an impaired maturation of muscle fibres, and caused by defects in the MTM1 gene. This gene defines a new family of putative tyrosine phosphatases conserved through evolution. We have determined intronic flanking sequences for all the 15 exons to facilitate the detection of mutations in patients and genetic counselling. We characterised a new polymorphic marker in the immediate vicinity of the gene, which might prove useful for linkage analysis. Sequencing of the TATA-less predicted promoter provides the basis for transcriptional regulatory studies.

Basis of phenotypic variability in sporadic Creutzfeldt-Jakob disease.

OBJECTIVE: To determine the correlation of clinical and pathologic features with prion protein (PrP) gene polymorphism at codon 129 and with biochemical characteristics of the protease-resistant PrP (PrPres) in sporadic Creutzfeldt-Jakob disease (CJD). METHODS: Clinical data acquisition, determination of the codon 129 genotype of the PrP gene, brain pathologic study, and immunoblot analysis of crude brain extracts were carried out in 14 patients. RESULTS: The first group of 10 subjects showed the classic clinical triad, with dementia, myoclonus, and periodic sharp waves on EEG. None of the subjects had amyloid plaques, but PrP immunoreactivity was of diffuse synaptic type in the cerebellar cortex. All subjects were methionine-methionine at codon 129 and the PrPres had a biochemical profile of type 1 (unglycosylated band of 21.5 kD). A second group of three patients showed cerebellar ataxia and later dementia. Periodic sharp waves on EEG were absent. PrP amyloid plaques predominated in the cerebellar cortex, along with diffuse PrP immunoreactivity. These subjects were valine-valine at codon 129 and had a type 2 PrPres (unglycosylated band of 19.4 kD). In the last patient cerebellar ataxia and dementia appeared simultaneously. Many Kuru-type plaques were present in the cerebellar cortex; many PrP amyloid plaques were present in the basal ganglia. This patient was methionine-valine at codon 129 and the PrPres was of type 1. CONCLUSIONS: The codon 129 genotype is only one of the factors determining CJD phenotype, and the biochemical pattern of PrP has no direct correlation with this phenotype.

Is clonidine growth hormone stimulation a good test to differentiate multiple system atrophy from idiopathic Parkinson's disease?

Clonidine, a centrally active alpha 2-adrenoreceptor agonist used to lower blood pressure, has been proposed to differentiate central from peripheral autonomic deficits and multiple system atrophy (MSA) from untreated idiopathic Parkinson's disease (IPD). A lack of growth hormone (GH) increase after clonidine infusion is found in patients with MSA, but not in those with IPD or with pure autonomic failure. We studied 19 IPD and 7 MSA patients to assess whether this test could be used in clinical practice to distinguish MSA from IPD, whatever the stage of the disease. Serum GH levels were measured 15, 30, 45 and 60 min after a 10-min infusion of 2 micrograms/kg clonidine. GH levels remained stable after clonidine infusion in all 7 MSA patients but increased in only 12 of the 19 IPD patients, while remaining stable in the other 7. No correlation was found with the presence of orthostatic hypotension. We conclude that the GH response to clonidine infusion has a very high sensitivity (100% in our series and in previous studies) for the diagnosis of MSA. However, this response cannot be used as a diagnostic test because of its poor specificity.

Bacterial keratitis: predisposing factors, clinical and microbiological review of 300 cases.

AIM: To identify predisposing factors and to define clinical and microbiological characteristics of bacterial keratitis in current practice. METHODS: A retrospective analysis of the hospital records of patients presenting with bacterial keratitis and treated at the Quinze-Vingts National Center of Ophthalmology, Paris, France, was performed during a 20 month period. A bacterial keratitis was defined as a suppurative corneal infiltrate and overlying epithelial defect associated with presence of bacteria on corneal scraping and/or that was cured with antibiotic therapy. Risk factors, clinical and microbiological data were collected. RESULTS: 300 cases (291 patients) of presumed bacterial keratitis were included. Potential predisposing factors, usually multiple, were identified in 90.6% of cases. Contact lens wear was the main risk factor (50.3%). Trauma or a history of keratopathy was found in 15% and 21% of cases, respectively. An organism was identified in 201 eyes (68%). 83% of the infections involved Gram positive bacteria, 17% involved Gram negative bacteria, and 2% were polymicrobial. Gram negative bacteria were associated with severe anterior chamber inflammation (p=0.004), as well as greater surface of infiltrates (p=0.01). 99% of ulcers resolved with treatment, but only 60% of patients had visual acuity better than the level at admission, and 5% had very poor visual outcome. CONCLUSIONS: Contact lens wear is the most important risk factor. Most community acquired bacterial ulcers resolve with appropriate treatment.

[Auditory P300 in subjects in a post-anoxic coma. Preliminary data]. / Etude du P300 auditif chez des sujets en coma post-anoxique. Données préliminaires.

The first results concerning the role of event related potentials (P300) in assessing a prognosis in comatose patients reported in the literature have been encouraging. The cause of the coma is an important prognostic factor by itself, especially when traumatic and anoxic comas are compared, with a less favorable prognosis in anoxic coma. In our study, only anoxic patients have been investigated using somatosensory evoked potentials, brainstem auditory evoked potentials P300 auditory event-related potentials. Clinical evaluation was performed using Glasgow, Liege and Jouvet coma scales. In twenty patients studied, six had a positive P300 and three of them awake. Out of the 14 patients without a P300 only one awoke (5%), 12 patients died and one is in deep coma. Despite the small number of patients in this study, the first results confirm the interest of auditory P300, in addition to clinical evaluation and evoked potential testing.

[Myotubular myopathy]. / La myopathie myotubulaire.

The myotubular myopathy is an X-linked centronuclear myopathy characterized by severe neonatal hypotonia and generalized muscle weakness which most frequently results in the premature death of the newborn infants by respiratory failure. The characteristic muscle histopathology consists in centrally positioned nuclei in most muscle fibers. In 1996, the gene responsible for the disease, MTM1, was identificated in Xq28 region. Since then, more than hundred mutations have been isolated. Genotype - phenotype correlation is complex because mutations are found along the entire coding sequence of the MTM1 gene. Most mutations are associated with very severe phenotype with death before the first year of life, however a milder phenotype has been individualized. It is important to be aware of the existence of such milder MTM phenotype because in those patients a very mild expression permit normal life into adulthood.

[Hemicontracture and facial myokimia as the first manifestation of multiple sclerosis]. / Hémicontracture et myokimies faciales continues révélatrices d'une sclérose en plaques.

Continuous facial myokimia (CFM) are characterized by a contracted facial appearance and vermicular movements that spread across a hemiface. Pontine lesions are their most frequent cause. Although their occurrence during the course of multiple sclerosis is not uncommun, CFM are rarely the first sign of the disease. We report two cases of CMF as first manifestation of multiple sclerosis. Multiple sclerosis is the first diagnosis to suspect in young people with CFM.

[Spinal cord MRI in multiple sclerosis: comparison of turbo-spin echo sequences and turbo-MRI]. / IRM médullaire dans la sclérose en plaques: comparaison des séquences turbo-spin écho et turbo-IRM.

UNLABELLED: To compare T2-weighted turbo spin-echo (turboSE) and turbo inversion recovery magnitude (turboIRM) sequences in the detection of multiple sclerosis of the spinal cord. METHODS: 28 patients with clinically confirmed multiple sclerosis and signs of cervical cord involvement were examined on a 1.5 tesla MR system. Sagittal T2-weighted turboSE sequences were followed by sagittal turboIRM sequences. 24 patients also underwent a cranial study. RESULTS: Using turboIRM sequence, all the patients showed at least one spinal cord plaque with a total of 91 plaques. Using turboSE sequence, 61 lesions were identified in 25 patients. Spinal plaques were seen best on turboIRM sequences with a higher contrast between the lesion and the normal spinal cord. In 10 patients with cord plaques in both sequences, the cranial study was normal or non conclusive. CONCLUSIONS: The turboIRM sequences detect more cord lesions and with a better contrast than the turboSE sequence. This results confirm those described in other studies and demonstrate the inadequacy of the fast spin echo sequences. Assessment of spinal plaques is particularly important when MR findings of the brain are negative or non conclusive.

[Measurement of total IgE in tears : the adaptation of an immunoenzyme technique and the value of investigating locally produced IgE in the diagnosis of chronic conjunctivitis]. / Dosage des IgE totales dans les larmes: adaptation d'une technique immuno-enzymatique et intéret de la recherce d'une syntheèse locale d'IgE dans l'exploration des conjonctivites chroniques.

The measurement of total IgE in tears could be useful to confirm the diagnosis of allergic conjunctivitis, particularly in chronic forms when the etiology is difficult to determine solely on clinical criteria and allergologic anamnesis. Since the level of IgE is normally very low in tears, the adaptation of a fluorescent enzyme immunoassay of total IgE (Unicap, Pharmacia-Upjohn) reduces the detection limit from 0.35 to 0.15 kU/l. Satisfactory results are obtained for accuracy, within-run precision (CV < 5%) and between-run precision (CV < or =11%). During allergic reactions, the local synthesis of IgE is evaluated by the ratio between the measured lacrimal IgE and the lacrimal IgE that have been filtered through the hemato-lacrimal barrier. The serum and tear albumin allows to quantify the permeability of the barrier and thus the amount of filtered tear IgE.

[Protein evaluation of tears: different biological parameters and their respective value]. / Le bilan protéique des larmes: les différents paramètres biologiques et leur intérêt respectif.

PURPOSE: The lacrimal film contains 6 to 10 g/l of proteins, 99% of which locally synthesized by the lacrimal glands. The study of these proteins allows us to explore the lacrimal function and to reveal an inflammatory process. MATERIAL AND METHODS: The lacrimal proteinic profile included the determination of total proteins and electrophoresis on agarose gel and, if necessary, specific determinations of albumin, lactoferrin, lysozyme and immunoglobulins using a nephelemetric technique. Normal values were established on a hundred of individual tears. RESULTS: The electrophoretic proteinic profile may present different abnormalities, such as an inflammatory process, a functional alteration of the lacrimal glands or a dysproteinic abnormality of the tears. The specific determination of the principal lacrimal proteins allows us to accurately quantify each of them. CONCLUSION: The electrophoresis of the tears on an agarose gel reveals the presence of an inflammatory process or the quantitative or qualitative alteration of the lacrimal function. The immuno-nephelemetric determination of the most important proteins which are involved in theses mechanisms gives an accurate quantitative measurement of proteins and allows biological follow-up of the disease.

[2-year retrospective study of 344 patients presenting for corneal lesion and who were examined for free amebas]. / Etude rétrospective sur deux ans de 344 patients ayant consulté pour une lésion cornéenne et chez lesquels une recherche d'amibes libres a été effectuée.

PURPOSE: The number of cases of amibian keratitis seems to have increased in the XV-XX hospital. Consequently, a study was carried out on 344 patients who came to be treated for keratitis or corneal ulcers over the last two years. METHODS: 28 patients out of 344 showed Acanthamoeba in lens storage cases and/or corneal scrapes. The diagnosis, treatment and clinical evolution of 28 patients were presented. RESULTS: 26/28 patients wore contact lenses, 22 lens cases examined out of 149 (15%) and 7 of corneal scrapes out of 344 (2%) showed the presence of Acanthamoeba. 68% of patients (19 out of 28) came to be treated for the first time in the emergency department. 2/28 patients (7%) were examined at the very beginning of the amibian infection and 24/28 patients (86%) showed the beginnings of stromal infiltration. The diagnosis for 13 of the patients was made within 15 days. 19/28 patients recovered, 1 patient had to undergo a penetrating keratoplasty, 4/28 patients had bacterial infections and 4/28 patients disappeared and we heard nothing more from them. CONCLUSION: Acanthamoeba were isolated from only 7 cornea, whereas 24 patients had an amibian infection. A deep corneal scrape is necessary to avoid a false negative result. A lens storage cases examination is highly recommended.