One-year prospective real-world assessment of effectiveness and safety of erenumab in migraine prevention: results of the French FHU INOVPAIN registry study.

BACKGROUND: Randomized clinical trials have demonstrated efficacy and safety of erenumab. The aim of this study is to evaluate the effectiveness and safety of erenumab in a real-world setting in French patients with migraine associated with extreme unmet needs. METHODS: This is a one year-prospective real-word study with enrolment of all consecutive adult patients included in the FHU InovPain registry who participated in a compassionate erenumab use program. RESULTS: Of 144 patients included, 140 patients (82.1% female / mean age of 50.9 ± 11.4) received at least one dose of erenumab and were concerned by effectiveness and safety assessment. All patients had failed 11 oral preventive treatments. Most of them suffered from chronic migraine (88.6%) and presented a medication overuse (90.7%) at baseline. Thirty-eight (27.1%) discontinued treatment during the 12-month follow-up, with 22 (15.7%), 11 (7.9%) and 5 (3.6%) patients before 3, 6 or 9 months of treatment. The proportion of ≥ 50% responders at M3, M6, M9 and M12 was 74/140 (52.9%), 69/118 (58.5%), 61/107 (57.0%) and 60/102 (58.8%) respectively. At M3, the rate of reversion from chronic migraine to episodic migraine was 57.3% and the rate of transition from medication overuse to non-overuse was 46.5%. For monthly migraine days, the median (IQR) was 18.0 (13.0-26.0), 9.0 (5.0-17.0), 7.5 (5.0-14.0), 8.0 (5.0-12.5) and 8.0 (5.0-12.0) at M0, M3, M6, M9 and M12 respectively. For HIT-6 score, the median (IQR) was 68.0 (63.8-73.3), 60.0 (54.0-65.0), 60.0 (50.3-53.0), 59.0 (50.0-63.0) and 58.0 (50.0-62.9) at M0, M3, M6, M9 and M12 respectively. Fifty-three (37.9%) patients reported at least one of the following adverse events: cutaneous erythema and/or pain at the injection site for 42 (30%) patients, constipation for 22 (15.7%) patients, muscle spasm for 2 (1.4%) patients, alopecia for one (0.7%) patient and blood pressure increase in one (0.7%) patient. There was no serious adverse event. One female patient became pregnant after 5 months of exposure to erenumab with a safe evolution after treatment discontinuation. CONCLUSION: This first French real-world study related to migraine prevention with CGRP-mAbs confirms effectiveness and safety of erenumab in patients with extreme unmet needs.

Temocillin against Enterobacteriaceae isolates from community-acquired urinary tract infections: low rate of resistance and good accuracy of routine susceptibility testing methods.

Background: Temocillin is an old 'revived' antibiotic that may play an important role in the treatment of febrile urinary tract infection (UTI). Data regarding its activity against current Enterobacteriaceae isolates as well as the performance of routine susceptibility testing methods are, however, scarce. Objectives: To determine the MICs of temocillin for Enterobacteriaceae strains reflecting the current epidemiology and to analyse the accuracy of three commercial methods. Methods: Enterobacteriaceae isolates causing community-acquired UTI were prospectively collected from September 2015 to January 2017 in two French centres. Temocillin MIC was determined by agar dilution (AD) as the reference method and then compared with: (i) susceptibility testing by disc diffusion; (ii) MIC determination by Etest; and (iii) MIC estimation by the Vitek 2 automated system. Results: A total of 762 Enterobacteriaceae were analysed comprising 658 (86.4%) Escherichia coli and 37 (4.9%) ESBL-producing isolates. Susceptibility rate assessed by AD was 99.6% according to the 8 mg/L clinical breakpoint and was significantly lower against the ESBL-producing isolates than the non-ESBL-producing isolates (94.6% versus 99.9%, P < 0.01). The MIC50 and MIC90 for the total set were 3 and 6 mg/L, respectively. According to the 8 mg/L clinical breakpoint, the major error rate was <1% for disc diffusion and Etest, and significantly higher for Vitek 2 (4.3%, P < 0.01), but still low. No very major error was noticed. Conclusions: Temocillin showed a high level of activity against Enterobacteriaceae from community-acquired UTI and good to excellent reliability of routine methods for susceptibility testing in such a setting.

Impact of improved urban environment and coaching on physical condition and quality of life in elderly women: a controlled study.

BACKGROUND: Physical activity can slow the ageing process and preserve autonomy in the elderly. The aim of this study was to assess the combined impact of an organized urban walking circuit and individual coaching on women senior citizens' physical well-being and quality of life. METHODS: Insufficiently physically active women >65 years were included in a quasi-experimental trial. Active arm: District with improved urban environment (IUE). Control arm: District without improved urban environment (WIUE). In each district, subjects were randomly allocated to receive coaching (C+ vs. C-). The main outcome measures were endurance, physical activity score, flexibility, quality of life, physical self-esteem, ageing exercise stereotypes, functional health and perceived health at baseline, three (M3) and six (M6) months. RESULTS: Fifty-two insufficiently physically active women were included, 23 in IUE and 29 in WIUE. Groups were comparable at baseline. At M3, endurance and physical activity score significantly improved compared with baseline in the IUE group and in the C+ group while no statistically significant change was observed for the WIUE group and the C- group. Moreover, endurance score was higher in the IUE group, whether coupled with coaching or not. After the coaching was removed, the IUE group regresses to baseline overall and the WIUE shows a decrement in endurance. CONCLUSION: Our study highlights the positive impact of an improved environment and of individual coaching on the level of physical activity and quality of life of insufficiently physically active elderly women.

A decreasing CD4/CD8 ratio over time and lower CSF-penetrating antiretroviral regimens are associated with a higher risk of neurocognitive deterioration, independently of viral replication.

Persistent immune activation is one of the suspected causes of HIV-associated neurocognitive disorders (HAND) in cART era. The CD4/CD8 ratio has been recently showed as a marker of immune activation and HAND. Our aim was to analyze if a decrease in the CD4/CD8 ratio over time could have an impact on neurocognitive deterioration. Randomly selected HIV-infected patients were followed for neuropsychological (NP) testing during a period of almost 2 years. Tests were adjusted for age, gender, and education. Patients were divided into 5 groups: normal tests (NT), neuropsychological deficit (ND, one impaired cognitive domain), asymptomatic neurocognitive disorders (ANI), mild neurocognitive disorders (MND), and HIV-associated dementia (HAD). Risk factors for neurocognitive deterioration were analyzed. Two hundred fifty-six patients underwent NP tests and 94 participated in the follow-up. The groups were comparable. Upon neuropsychological re-testing, six patients showed clinical improvement, 30 had worsened, and 58 were stable, resulting in 42 patients presenting with HAND (45 %). The majority of HAND cases consisted of ANI (26 %) and MND (16 %). In patients whose NP performance worsened, CPE 2010 score was lower at inclusion (7.13 vs 8.00, p = 0.003) and CD4/CD8 decrease more frequent (60 vs 31 %, p = 0.008) than in those who were stable or improved. Multivariate analysis confirmed these results. A decreasing CD4/CD8 ratio during a longitudinal follow-up of randomly selected HIV-infected patients and lower CSF-penetrating regimens were independently associated with cognitive decline. Monitoring trends in CD4/CD8 ratio could contribute to identifying patients at higher risk of neurocognitive deterioration.

Virologically suppressed patients with asymptomatic and symptomatic HIV-associated neurocognitive disorders do not display the same pattern of immune activation.

OBJECTIVES: Inversion of the CD4:CD8 ratio is a marker of immune activation and age-associated disease. We measured the CD4:CD8 ratio as a marker of cognitive impairment in HIV-infected patients and explored differences according to clinical severity. METHODS: Post hoc analysis of data from two prospective cohorts of HIV-infected patients randomly selected to undergo neuropsychological tests was performed. Test scores were adjusted for age, gender and education. Inclusion criteria were undetectable viral load and stable treatment for at least 6 months. Subjects with HIV-associated dementia were excluded. Patients were divided into an unimpaired group, a group with asymptomatic neurocognitive disorder (ANI) and a group with symptomatic HIV-associated neurocognitive disorder (sHAND), represented by mild neurocognitive disorder (MND). Demographic and background parameters, immune activation markers and the CD4:CD8 ratio were recorded. RESULTS: Two hundred patients were included in the study. The mean age was 52 years, 78% were male, the mean CD4 count was 624 cells/µL, the mean nadir CD4 count was 240 cells/µL, 27% were hepatitis C virus (HCV)-coinfected, the mean duration of HIV infection was 16 years, and the mean time on current combination antiretroviral therapy (cART) was 2.9 years. Twenty-nine per cent of subjects had HAND (21% had ANI and 8% had MND). In multivariate analysis, a CD4:CD8 ratio < 1 was associated with a nadir CD4 count < 200 cells/µL [odds ratio (OR) 3.68] and with the presence of CD4(+) CD38(+) HLA(+) cells (OR 1.23). Multinominal logistic regression showed that, in comparison with the unimpaired group, diagnosis of sHAND was associated with a CD4:CD8 ratio < 1 (OR 10.62), longer HIV infection (OR 1.15) and longer current cART (OR 1.34), while the ANI group differed from the unimpaired group only for education level. CONCLUSIONS: Aviraemic patients with sHAND did not display the same pattern of immune activation as subjects with ANI, suggesting that the underlying pathophysiological mechanisms could be different.

[Staphylococcus saprophyticus isolated from urine culture in outpatients: epidemiology and antimicrobial susceptibility (Label Bio Elbeuf study - November 2007-July 2009)]. / Staphylococcus saprophyticus isolés d'examens cytobactériologiques urinaires en ville : épidémiologie et sensibilité aux antibiotiques (étude Label Bio Elbeuf - novembre 2007-juillet 2009).

OBJECTIVE: The aim of the study was to determine the distribution and the antibiotic susceptibility of Staphylococcus saprophyticus patterns isolated from urine culture in outpatients (population: 57,000, Elbeuf, Normandie, France). DESIGN: Prospective study from November 2007 to October 2009 in collaboration with three private medical laboratories. Determination of susceptibility to oxacillin by disk diffusion (cefoxitin, and moxalactam), automated method (Vitek BioMérieux 2) and mecA PCR's detection. Determination of the minimum inhibitory concentration by microbroth dilution for other antibiotics. RESULTS: Five thousand and fifty-one bacterial strains isolated, 91 strains of S. saprophyticus (1.8%), 89 in women (2.25%) and two in men (0.18%). S. saprophyticus represented 10.3% and 14.5% of isolates (women respectively aged between 11 and 30; 16 and 20 years); S. saprophyticus is isolated less frequently in winter. mecA PCR detection was positive for two strains. All strains tested were susceptible to ciprofloxacin and furans. Only one strain is resistant to cotrimoxazole. CONCLUSIONS: S. saprophyticus is found mostly in women between 11 to 30 years. Cotrimoxazole (after susceptibility testing) is efficient in case of S. saprophyticus's cystitis. Furans (probabilistic treatment) have to be reevaluated because of the potential for serious adverse effects.

[Lactobacillus delbrueckii: probable agent of urinary tract infections in very old women]. / Lactobacillus delbrueckii : probable cause d'infections urinaires chez la femme très âgée.

Lactobacillus urinary tract infection (UTI) seems exceptionally reported. Nevertheless, with the introduction of a chromogenic medium UriSelect 4, eight cases of UTI in old women (mean of 81.2 years) mediated by Lactobacillus delbrueckii identified by DNA sequencing were reported between 2007 and 2009.

The course of primary progressive aphasia diagnosis: a cross-sectional study.

BACKGROUND: The primary progressive aphasia (PPA) diagnosis trajectory is debated, as several changes in diagnosis occur during PPA course, due to phenotype evolution from isolated language alterations to global cognitive impairment. The goal of the present study, based on a French cohort, was to describe the demographics and the evolution of subjects with (PPA) in comparison with Alzheimer's disease (AD) on a period of 7 years. METHODS: We conducted a repeated cross-sectional study. The study population comprised individuals with PPA and AD diagnosis (N=167,191) from 2010 to 2016 in the French National data Bank (BNA). Demographic variables, MMSE scores, diagnosis status at each visit and prescribed treatments were considered. RESULTS: From 2010 to 2016, 5186 patients were initially diagnosed with PPA, 162,005 with AD. Compared to AD subjects, significant differences were found concerning age (younger at first diagnosis for PPA), gender (more balanced in PPA), education level (higher in PPA) and MMSE score (higher of 1 point in PPA). Percentage of pending diagnosis, delay between first consultation and first diagnosis and the number of different diagnoses before the diagnosis of interest were significantly higher in PPA group compared to AD group. Pharmacological and non-pharmacological treatments were significatively more recommended following PPA than AD diagnosis. CONCLUSION: This study improves the knowledge of PPA epidemiology and has the potential to help adopting appropriate public health service policies. It supports the hypothesis that PPA is diagnosed later than AD. The PPA diagnosis increases the prescription of non-pharmacological treatments, especially speech and language therapy (SLT) that is the main treatment available and most effective when at the initial stage. TRIAL REGISTRATION: ClinicalTrials.gov identifier NCT03687112.

Genetic diversity of meat quality related genes in Argentinean pigs.

Genetic influence on pork quality exists between breeds and within a breed. The variation is caused by a large set of genes, and pork quality traits have a multifactorial background. Research into the genetics of meat quality found causative mutations associated with marked effects on pig meat value. This study aimed to investigate the segregation of meat quality-related SNPs and compare their diversity and genetics in commercial and Creole pigs from different farms in the North-West of Argentina. A screen for SNPs in RYR1, PRKAG3, CAST, and SOX6 candidate genes and the differentiation of their genotypes by PCR-RFLP was conducted. All genes were characterized by a high level of polymorphism and heterozygosity, and populations showed no differences in the genetic structure for the analyzed SNPs. These results highlighted the role of pig genotypes as a source of basic variability potentially affecting processed meat products and fresh meat.

Prevalence of mycoplasma-like lung lesions in pigs from commercial farms from Spain and Portugal.

BACKGROUND: Mycoplasma hyopneumoniae causes a chronic respiratory disease that produces important economic losses due to poor productive performance, increased mortality and costs for several control strategies. The prevalence of mycoplasma-like lesions (MLL) at abattoir has been widely studied in different countries, making use of different scoring systems. However, most of them are difficult to apply in abattoirs with high number of pigs sacrificed per hour. For that reason, it is necessary to adapt the scoring system to the reality of the modern abattoir, even if there is a loss of accuracy. Our purpose was to investigate the prevalence and severity of MLL at abattoirs in Spain and Portugal using a 0 to 5 scoring system adapted to abattoirs with high number of sacrificed pigs per hour and to highlight the histopathological diagnosis as confirmatory method to identify patterns of pneumonia correlated to gross lesions. RESULTS: Cranioventral pulmonary consolidation, a typical MLL, was the most frequent lung lesion (30.97 %) detected at the abattoir, followed by dorsocaudal infarcts with pleurisy (12.51 %) and pleurisy alone (6.26 %). The average score for all examined lungs at abattoir was 1.99 out of 5 points. The histopathological study revealed that the 78.17 % of the randomly selected lungs with MLL presented microscopic lesions compatible with M. hyopneumoniae infection. Most bronchointerstitial and interstitial pneumonia lesions had a chronic course while most suppurative and fibrinous bronchopneumonia lesions had an acute course and a higher degree of severity. The combination of microscopic lesions more frequently observed was bronchointerstitial pneumonia + interstitial pneumonia + suppurative bronchopneumonia. CONCLUSIONS: The prevalence of MLL at abattoir was 30.97 %, however, after microscopic examination the real prevalence of lungs with lesions compatible with M. hyopneumoniae infection was reduced up to 24.21 %. The six more prevalent combinations of lesions in the microscopic study involved the 66.13 % of examined lungs, and in all of them, microscopic lesions characteristic of M. hyopneumoniae infection were found, what supports the importance of M. hyopneumoniae as a primary pathogen in cases of PRDC.

PCR-based methods to the diagnosis of imported malaria.

Rapid and precise diagnosis of malaria is needed to take care febrile patient returning from endemic areas. Since the first description of the diagnosis of Plasmodium infection by polymerase-chain-reaction (PCR), the role of this kind of molecular method in the laboratory diagnosis of imported malaria is still a topical question. PCR-based assays were found to be more sensitive and more specific than all conventional methods. The highest contribution of the molecular diagnosis is that a PCR negative result would ascertain the lack of any malaria infection, thus quickly orienting the investigations toward other aetiology. This technique should be now considered as the gold standard for the diagnosis of imported malaria.

Sex ratio in dementia with Lewy bodies balanced between Alzheimer's disease and Parkinson's disease dementia: a cross-sectional study.

BACKGROUND: Gender distribution varies across neurodegenerative disorders, with, traditionally, a higher female frequency reported in Alzheimer's disease (AD) and a higher male frequency in Parkinson's disease (PD). Conflicting results on gender distribution are reported concerning dementia with Lewy bodies (DLB), usually considered as an intermediate disease between AD and PD. The aim of the present study was to investigate gender differences in DLB in French specialized memory settings using data from the French national database spanning from 2010 to 2015 and to compare sex ratio in DLB with that in AD, Parkinson's disease dementia (PDD), and PD. Our hypothesis was that there is a balanced sex ratio in DLB, different from that found in AD and PD. METHODS: We conducted a repeated cross-sectional study. The study population comprised individuals with a DLB, AD, PDD, or PD diagnosis according to the International Classification of Diseases, Tenth Revision, in the French National Alzheimer Database between 2010 and 2015. Sex ratio and demographic data were compared using multinomial logistic regression and a Bayesian statistical model. RESULTS: From 2010 to 2015 in French specialized memory settings, sex ratios (female percent/male percent) were found as follows: 1.21 (54.7%/45.3%) for DLB (n = 10,309), 2.34 (70.1%/29.9%) for AD (n = 135,664), 0.76 (43.1%/56.9%) for PD (n = 8744), and 0.83 (45.4%/54.6%) for PDD (n = 3198). Significant differences were found between each group, but not between PDD and PD, which had a similar sex ratio. CONCLUSIONS: This large-sample prevalence study confirms the balanced gender distribution in the DLB population compared with AD and PD-PDD. Gender distribution and general demographic characteristics differed between DLB and PDD. This is consistent with the hypothesis that DLB is a distinct disease with characteristics intermediate between AD and PD, as well as with the hypothesis that DLB could have at least partially distinct neuropathological correlates.

[Campylobacter fetus bacteremia and cellulitis complicating a venous access port infection in an HIV infected patient]. / Bactériémie et dermohypodermite à Campylobacter fetus sur cathéter à chambre implantable au cours d'une infection par le VIH.

Campylobacter fetus subspecies fetus is an opportunist Gram-negative bacillus, which is known to be a cause of systemic infections, mainly in immunocompromised patients. We report a C. fetus bacteremia and cellulitis complicating a venous access port infection in a patient with acquired immunodeficiency syndrome (AIDS). This bacillus seems to have a predilection for the vascular endothelium and its isolation is difficult. Physicians should be aware of C. fetus infection in patients with vascular devices. Microbiologists should accurately isolate this organism from clinical specimens by modifying incubation techniques and performing molecular biology. The prognosis seems to be improved by a prolonged betalactam antibiotic regimen, especially amoxicilline plus clavulanic acid. In HIV infected patients, quinolones that were successful in our case, should be used with caution because of increasing resistance to antibiotics.

[Community-acquired urinary tract infections in 15 to 65 years old female patients in France. Susceptibility of E. coli according to history: AFORCOPI-BIO network 2003]. / Infections urinaires communautaires de la femme de 15 à 65 ans: sensibilité aux antibiotiques de E. coli en fonction des antécédents: étude AFORCOPI-BIO 2003.

OBJECTIVES: A multicenter study was implemented in order to determine the distribution and antibiotic susceptibility patterns of strains isolated from 15 to 65 year old female patients with community-acquired urinary tract infections. PATIENTS AND METHODS: From October to December 2003, 11 French private laboratories consecutively collected 420 clinical strains with medical data. Minimal inhibitory concentrations of antibiotics on E. coli were determined using the agar dilution method in a coordinating center and interpretation followed the recommendations of the Comité de l'antibiogramme de la Société française de microbiologie. RESULTS: Escherichia coli was the most prevalent pathogen (80%) followed by Proteus mirabilis (4%), Klebsiella spp (2%), other Enterobacteriaceae (4%), Enterococcus spp (3%), Staphylococcus aureus (2%), Staphylococcus saprophyticus (2%), and Streptococcus agalactiae (2%). The susceptibility of E. coli strains was 61% for amoxicillin (AMX), 93% for nalidixic acid (NAL), 97% for norfloxacin (NOR) and ciprofloxacin (CIP), 77% for cotrimoxazole (SXT), 99% for fosfomycin, gentamicin and cefotaxime. The susceptibility of E. coli was lower in case of previous treatment with beta-lactam antibiotics for AMX (84 vs 95% p=0.02) and SXT (62 vs 81% p=0.02). In the same way, previous treatment with quinolones was associated with decreased susceptibility for NAL (84 vs 95% p=0.02) and SXT (62 vs 81% p=0.02). CONCLUSIONS: In 2003, fluoroquinolones, third generation cephalosporins, aminoglycosides, and fosfomycin kept a good activity on E. coli collected from community-acquired urinary tract infections in 15 to 65 years old female patients in France.

Expression and purification of rotavirus proteins NSP5 and NSP6 in Escherichia coli.

Rotaviruses are one of the worldwide leading causes of gastroenteritis in children under 5 yr old. The rotavirus nonstructural NSP5 is a phosphoprotein implicated in viroplasms formation, whereas NSP6 could have a possible regulatory role of NSP5. It has been reported that N- and C-termini of NSP5 are important for their function. However, no structural information on NSP5 and NSP6 proteins is available. Because a high amount of protein is required for structural analysis, efficient expression systems are required. His-tag fusion at the C-terminus and glutathione-S-transferase (GST)-fusion at the N-terminus were used as expression systems, and conditions for recombinant proteins expression were obtained. His-tag fusion was not efficient to produce NSP5 (2% of total protein), but NSP6 was expressed in higher amounts (11% of total protein). In contrast, GSTNSP5 and GST-NSP6 proteins correspond to 34 and 31% of the total proteins, respectively. GST-fusions seem to have a protective effect against nonstructural rotavirus protein toxicity in Escherichia coli; however, in both systems, NSP5 and NSP6 recombinant proteins were expressed as inclusion bodies. Conditions for solubilization and purification of recombinant proteins were achieved. This is the first report of expression and purification of NSP5 and NSP6 recombinant proteins in suitable amounts for further structural analysis.

Prevalence of Plasmodium falciparum cytochrome b gene mutations in isolates imported from Africa, and implications for atovaquone resistance.

The atovaquone resistance of malaria parasites correlates with mutations in the cytochrome b gene. We sequenced the Plasmodium falciparum cytochrome b gene of 135 African isolates. Our data showed a high mutation rate (8.9%); however, the risk of emergence spreading of atovaquone-resistant P. falciparum strains could be limited.

Physicochemical and sensory properties of chicken nuggets with washed mechanically deboned chicken meat: Research note.

The effects of different proportions (0%, 10%, 20%, 30%, 40%) of washed mechanically deboned chicken meat (WM) as a substitute for hand deboned chicken meat, on the physicochemical and sensory characteristics of chicken nuggets were evaluated. The addition of WM increased fat content, but it was only significant (P<0.05) when 40% of WM was added, whereas the protein content was significantly (P<0.05) reduced as from 20%. Significant differences (P<0.05) were found in L(*), a(*) and b(*) values with different proportions of WM, however, these differences were evidently not discerned as shown by the no significant differences (P>0.05) in ΔE(*) color scores. The addition of WM did not affect (P>0.05) sensory attributes of chicken nuggets. From a technical viewpoint, up to 40% WM could be incorporated into nugget formulation instead of hand deboned chicken meat without affecting sensory attributes of the product. Minor changes in composition were observed but they were probably not detrimental to the product.

[A case of acquired angioedema]. / Un cas d'oedème angioneurotique acquis.

We report the case of a 46 year-old woman presenting an acquired angioedema. Angioedema is an C1 inhibitor deficiency. Patients present recurrent non inflammatory swelling of the head and extremities and recurrent attacks of severe abdominal pain. This clinical presentation is non specific : investigation of complement is useful for diagnosis. Laboratory testing show low serum levels of C4 with normal levels of C3. Low C1 esterase inhibitor confirm the diagnosis. If acquired angioedema, a cause must be searched.

[Utility and limitations of laboratory diagnosis of amebiasis]. / Intérêt et limites du diagnostic au laboratoire de l'amibiase en 2004.

Entamoeba histolytica is an invasive and pathogenic protozoan parasite that causes amebiasis. It must be distinguished from Entamoeba dispar, a nonpathogenic commensal parasite of the human gut lumen that is morphologically identical to Entamoeba histolytica. Diagnosis of amoebic colitis currently requires combination of microscopic examination of stool specimens with another technique allowing positive identification of the two species. Stool culture followed by zymodem analysis is considered as gold standard but is not applicable in routine practice. Detection of specific Entamoeba histolytica antigens in stools is a fast, sensitive technique that should be considered as the method of choice. Stool PCR is a highly sensitive and specific technique but high cost make it unsuitable for use in endemic areas where economic conditions are difficult. The utility of serologic tests in distinguishing Entamoeba dispar from Entamoeba histolytica is controversial. However serology is still considered as the method of choice for diagnosis of extraintestinal amebiasis. Circulating Gal/GalNac lectin antigens can be detected in the serum of 96% of patients with untreated amoebic liver abscess. In the future this method should allow early diagnosis and treatment of extraintestinal amoebiasis in patients who have not yet developed detectable serum antibodies.

Contribution of PCR-based methods to diagnosis and management of imported malaria.

Since the first description, in 1990, of the diagnosis of Plasmodium falciparum infection by polymerase-chain-reaction (PCR), the role of this kind of molecular method in laboratory diagnosis of imported malaria is still a topical question. Various molecular assays have been used, the first of which was hybridization using labeled probes in 1984. When compared to thick blood smear, this test displayed a sensitivity ranging from 65% to 81% and specificity was close to 100%. The next technical improvement was the introduction of the so-called polymerase chain reaction (PCR), the principle of which was described in 1985. In 1993, a PCR-based assay detecting all four Plasmodium species was published, followed by different variants of this method. By the turn of the century, novel real-time PCR slashed workaround time, which dropped from 2 1/2 hours to less than 1 hour. Moreover, automatic reading with no human action on PCR products reduced the risks of contamination. The first application of real-time PCR to the diagnosis of malaria was published in 2001. PCR-based assays were found to be more sensitive than all conventional methods. Variations in sensitivity were probably due to different medical practices as well as to the proportion of various types of subjects (travelers under chemoprophylaxis, immigrants from malaria-endemic areas) in the population undergoing malaria diagnosis. The target of the primers was also of crucial importance: for the detection of P. falciparum, the most efficient assays amplified either the gene SSUrRNA, or Pf155/RESA, or Cox 1. Specificity of PCR results is guaranteed by the nature of the target for primers or probes, as determined by the studies of the Plasmodium genome whose results are available in GenBank. PCR use often corrected the results of Plasmodium species identification by microscopy and PCR-based methods were found to be the most efficient for the detection of mixed infections. Concerning the diagnosis of imported malaria, it appears clearly that PCR should be considered as second-line method which can be especially interesting, as a negative result rules out malaria in febrile patients. However, the use of PCR assays appears to be restricted to health centers, such as University Hospitals, for whom malaria identification is an important and routine problem. In the future, the detection of mutations related to drug resistance could be used to orient anti-malarial therapy.