[Utility of real-time PCR for the diagnosis of infantile visceral leishmaniasis in Algeria]. / Contribution de la PCR en temps réel au diagnostic de la leishmaniose viscérale infantile en Algérie.

Visceral leishmaniasis (VL) is a parasitic disease and serious public health problem. The most severe form of all leishmaniases, it is fatal if untreated. Currently, it affects close to 540 people per year in Algeria, mainly in Kabylie, where it is fatal in 6% of all cases. This disease primarily affects children. Its diagnosis is based principally on visualization of parasites in bone marrow aspirate. Serologic studies are also used to diagnose VL, as is molecular biology, which has been demonstrated to be both faster and more sensitive. The aim of our study was to evaluate the utility of real-time polymerase chain reaction (RT-PCR) in the diagnosis of VL in endemic areas. Of the 108 samples analyzed, 32 were positive according to RT-PCR (29.6%); microscopy yielded positive result in 24% and serology in 25.9%. RT-PCR increased the overall number of diagnoses detected from 26 to 32, a 20% improvement over microscopic methods. In view of the lethal consequences of failure to diagnose this disease, any improvement in diagnostic methods provides direct medical benefit.

Invasive aspergillosis: an important risk factor on the short- and long-term survival of acute myeloid leukemia (AML) patients.

Invasive aspergillosis (IA) during induction chemotherapy of acute myeloid leukemia (AML) could worsen the prognosis. Our objective was to study how the development of IA during AML interferes with the therapeutic strategy and to evaluate its impact on the short- and long-term survival. Newly diagnosed AML patients between the years 2004 and 2007 were retrospectively analyzed. The outcome was death of the patient. A Cox proportional hazards model with the diagnosis of IA and post-induction response evaluation as the main exposure was fitted. Overall, 262 patients were analyzed and 58 IA were observed. The 2-year survival of patients having had remission of AML was 54% and, for patients with failure of chemotherapy, it was 5% (p < 0.001). The 2-year survival of patients having had IA was 14%, and without IA, it was 32% (p = 0.01). Multivariate analysis showed that IA was associated with a higher risk of death in case of remission compared to no IA (hazard ratio [HR] = 1.66 [1.05-2.65], p = 0.031) and also in case of failure (HR = 6.43, p < 0.001). IA was associated with an increased risk of death for patients if they were either in remission or in failure after induction chemotherapy.

Epidemiological trends in invasive aspergillosis in France: the SAIF network (2005-2007).

A prospective (2005-2007) hospital-based multicentre surveillance of EORTC/MSG-proven or probable invasive aspergillosis (IA) cases whatever the underlying diseases was implemented in 12 French academic hospitals. Admissions per hospital and transplantation procedures were obtained. Cox regression models were used to determine risk factors associated with the 12-week overall mortality. With 424 case-patients included, the median incidence/hospital was 0.271/10(3) admissions (range 0.072-0.910) without significant alteration of incidence and seasonality over time. Among the 393 adults (62% men, 56 years (16-84 years)), 15% had proven IA, 78% haematological conditions, and 92.9% had lung involvement. Acute leukaemia (34.6%) and allogeneic stem cell transplantation (21.4%) were major host factors, together with chronic lymphoproliferative disorders (21.6%), which emerged as a new high-risk group. The other risk host factors consisted of solid organ transplantation (8.7%), solid tumours (4.3%), systemic inflammatory diseases (4.6%) and chronic respiratory diseases (2.3%). Serum galactomannan tests were more often positive (≥69%) for acute leukaemia and allogeneic stem cell transplantation than for the others (<42%; p <10(-3)). When positive (n = 245), cultures mainly yielded Aspergillus fumigatus (79.7%). First-line antifungal therapy consisted of voriconazole, caspofungin, lipid formulations of amphotericin, or any combination therapy (52%, 14%, 8% and 19.9%, respectively). Twelve-week overall mortality was 44.8% (95% CI, 39.8-50.0); it was 41% when first-line therapy included voriconazole and 60% otherwise (p <0.001). Independent factors for 12-week mortality were older age, positivity for both culture and galactomannan and central nervous system or pleural involvement, while any strategy containing voriconazole was protective.

[Disseminated strongyloidiasis with parasitemia in a patient under corticosteroid-treatment]. / Anguillulose disséminée avec parasitémie chez un patient sous corticothérapie.

We report a case of disseminated strongyloidiasis with parasitemia in a 53-year-old man under corticosteroid therapy. It occurred more than 16years after contamination and led to severe sepsis and acute respiratory failure with P. aeruginosa pneumonia. The patient recovered after specific treatment (thiabendazole followed by albendazole) and antibacterial drugs.

[New use of primaquine for malaria]. / Le nouvel âge de la primaquine contre le paludisme.

Primaquine is the only available drug to treat Plasmodium vivax liver stages (hypnozoites). It has been used for more than five decades and is now included in an increasing number of clinical guidelines. The major concern is induced hemolysis when administered to glucose-6-phosphate-dehydrogenase deficient patients. Primaquine could be used for causal prophylaxis during and after exposure or for presumptive antirelapse therapy (PART) in case of high exposure to P. vivax. A radical cure is used to avoid relapse for patients with a confirmed bloodstream infection with P. vivax or P. ovale. In France, primaquine is not approved for prevention and treatment and its use requires a specific temporary authorization.

[Outsourced real-time PCR diagnosis of cutaneous leishmaniasis in the outbreak region of Constatine, Algeria]. / Diagnostic delocalise par PCR temps réel de la leishmaniose cutanée sévissant dans le foyer de Constantine (Algérie).

Taking into account the re-emergence of leishmaniasis in the world, the geographic variability of its epidemiology and the growing numbers of travellers, a pilot study on the diagnosis of cutaneous leishmaniasis was undertaken in Constatine, one of the outbreak regions in eastern Algeria. A total of 143 specimens were collected on blotters and tested by real-time PCR. Results were compared with those of direct examination. Diagnosis was positive for leishmaniasis in 81% of cases using PCR versus 48% of cases using microscopy. Real-time PCR showed a significant quantitative difference between patients for whom microscopic diagnosis was positive and those for whom direct examination was negative. The results presented in this study demonstrated the effectiveness and sensitivity of PCR in the diagnosis of cutaneous leishmaniasis from blotter specimens. This technique enabled in-field collection of specimens from each patient and provided prompt results. North-South cooperation based on the use of simple means for transmission of specimens for molecular diagnosis allowed creation of an effective partnership for daily diagnosis and promoted exchange between investigators in preparation for technology transfer.

Absence of nucleotide polymorphism in a Plasmodium vivax multidrug resistance gene after failure of mefloquine prophylaxis in French Guyana.

Vivax malaria is widespread and resistance has been described for chloroquine and sulfadoxine-pyrimethamine. We report on evidence of failure of mefloquine prophylaxis in a French soldier who contracted Plasmodium vivax in French Guyana, South America. Despite regular weekly mefloquine prophylaxis (250 mg/d), the patient presented with a first episode of vivax malaria, which was treated by chloroquine alone, then experienced a second crisis in France. The reappearance of the parasites occurred one day after the end of prophylaxis, confirming parasitological and clinical resistance in a non-immune patient. Mefloquine was detected by a liquid chromatography assay in plasma at a level of 1062 ng/ml, which was higher than the expected concentration after five months of weekly prophylaxis. This isolate had no single nucleotide polymorphisms of the pvmdr1 gene at seven allele positions: pvmdr1 N91, Y189, Y976, S1071, F1076, N1079 and D1291, corresponding to codons 86, 184, 939, 1034, 1039, 1042 and 1246 in P. falciparum. This observation of failure of mefloquine prophylaxis against P. vivax, when added to previously reported chloroquine and atovaquone-proguanil failure, strengthens the case for re-evaluating drug policies for vivax malaria and the need for continuous research on molecular markers of drug resistance.

Comparative diagnostic performance of two commercial rapid tests for malaria in a non-endemic area.

In the study reported here, the diagnostic performance of two new rapid tests for the diagnosis of malaria was evaluated in symptomatic patients in a non-endemic area. Of 557 consecutive patients, 109 (19.6%) had documented malaria. For the NOW ICT MALARIA P.f./P.v. (Binax, Portland, ME, USA) and OptiMAL IT (Diamed, Cressier, Switzerland) tests, respectively, sensitivity values were 96.3% and 79.8% (P-value, 0.0001), and specificity values were 98.8% and 98.4%. The NOW ICT test did not detect two of 80 Plasmodium falciparum infections, and it generated false-positive results for five patients. The OptiMAL IT test failed to detect ten of the P. falciparum infections, and it generated seven false-positive results. The results suggest that these rapid diagnostic tests for malaria may be useful, but they cannot replace microscopic examination of blood films.

Severe peritonitis due to Balantidium coli acquired in France.

The case reported here concerns an alcoholic pork-butcher who presented with severe colitis with peritonitis, caused by the only ciliate protozoan capable of infecting humans, Balantidium coli. This parasite is common in a variety of domestic and wild mammals, mainly pigs; however, its prevalence rate in humans is very low--particularly in industrialised, northern countries, including France. The infection is most frequently acquired by ingesting food or water contaminated by pig faeces, and it may be asymptomatic or may cause acute diarrhoea. Specific antibiotic treatment is efficacious, and it is important to consider the risk of this parasitic disease in susceptible patients presenting with bloody diarrhoea.

[Real-time PCR for detection of molecular markers of resistance in Plasmodium falciparum]. / Détection des marqueurs moléculaires de la résistance de Plasmodium falciparum par PCR en temps réel.

Plasmodium falciparum drug resistance is a major problem in malaria endemic areas. Molecular markers and in vitro tests have been developed to study and monitor drug resistance. However, none used alone, can provide sufficient data concerning the level of drug resistance and to issue precise guideline for drug use policies in endemic areas. We propose real-time PCR for the simultaneous detection of pfcrt and pfmdr1 genes mutations. The aim of this study was not to provide definitive data concerning the rate of mutations in an endemic area, but to describe a powerful method allowing the detection of major pfmdr1 and pfcrt mutations.