Pediatric visceral leishmaniasis diagnosis in Tunisia: comparative study between optimised PCR assays and parasitological methods.

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.

[Antiphospholipid antibodies in 146 women with repeated pregnancy losses]. / Anticorps antiphospholipides et pertes foetales: étude rétrospective de 146 cas.

Antiphospholipid antibodies are associated with arterial and venous thrombosis and recurrent abortions. However, the prevalence of these antibodies in repeated miscarriages varies in different reports. To obtain quantitative data with restricted criteria and discuss the origin of the variability on the literature, we investigated the presence of antiphospholipid antibodies in 146 women who had 2 or more consecutive pregnancy losses and in 99 women whose pregnancies were successful. Antiphospholipid antibodies (lupus anti-coagulant or anticardiolipin antibodies of 20 or more IgG units) were found in 45% of women with pregnancy losses and in 9% of controls (p < 0.001). The type of loss was determined according to the trimester of pregnancy and the time of the fetal loss. 68% of patients with antiphospholipid antibodies had at least one fetal loss on the second or third trimester compared with 45% of patients without fetal loss (p < 0.01). Further studies should be conducted using more rigorous definition of clinical and laboratory characteristics in a way to allow better comparison between studies.

[Peripheral blood culture as a diagnostic modality for visceral leishmaniasis: apropos of 61 cases]. / La culture du sang peripherique comme moyen diagnostique de la Leishmaniose viscerale: a propos de 61 cas.

The purpose of this prospective study was to assess the efficacy of culture of peripheral blood samples on NNN medium for detecting Leishmania. A total of 61 peripheral venous blood samples were tested. Most samples were collected from immunocompetent children presenting visceral leishmaniasis. The sensitivity of peripheral blood culture was 61%. Concordance with conventional bone marrow culture was good but the marrow technique was quicker. Since it is minimally invasive and easy to perform, peripheral blood culture is an attractive diagnostic alternative in immunocompetent patients. In addition peripheral blood culture presents a major epidemiological advantage in allowing identification of the causative strain of Leishmania if bone marrow culture was not performed for isoenzymatic characterization.

Visceral leishmaniasis in Tunisia: result of the isoenzymatic characterization of 65 Leishmania infantum strains.

The isoenzymatic characterization of 65 Tunisian strains of Leishmania, isolated from human cases of visceral leishmaniasis between June 1998 and August 2001, revealed the existence of 3 zymodemes of the L. infantum complex: MON-1 the most common (93.8%), followed by MON-24 (3.1%) and MON-80 (3.1%). 72% of the strains were obtained from children under the age of 5 years. The majority of the studied strains originated from 8 provinces of northern Tunisia, particularly the province of Zaghouan.

[Infantile visceral leishmaniasis from Leishmania infantum MON-24: a reality in Tunisia]. / Leishmaniose viscérale infantile à Leishmania infantum MON-24: une réalité en Tunisie.

The authors report the first documented observation of infantile visceral leishmaniasis due to Leishmania infantum MON-24 zymodeme in Tunisia. This zymodeme was isolated from a non-immunodepressed two-year-old child, with visceral leishmaniasis, originating from Zaghouan, in northern Tunisia. This case shows that beside L. infantum MON-1 and MON-80, L. infantum MON-24, the usual parasite of cutaneous leishmaniasis, there exists also a causative agent of visceral leishmaniasis in our country.

[Screening and confirmation of anti-HCV antibodies in Tunisian blood donors]. / Dépistage et confirmation des anticorps anti-VHC chez les donneurs de sang tunisiens.

Antibodies to Hepatitis C virus (HCV) were tested in 43000 Tunisian blood donors by using enzyme immuno-assay. Our results show that 0.7% (304/43000) were anti-HVC positive. Of these 304.78 were confirmed anti-HCV positive (0.18%) by immuno-blot, and 99 displayed an indeterminate profile. Different immune responses were observed: In donors with positive serologic pattern (78/304), 25.6% response towards whole antigens (C + NS3 + NS4 + NS5) was frequently observed (44/78) 56.4%. Reactivity to 2 antigens was observed in 28.2% (22/78) and with 3 antigens in only 15.4% (12/78), with systematic reactivity to core. In donors with indeterminate serologic pattern (99/304) 32.5%, reactivity to non-structural antigen NS5 was the most frequently observed (54/99) 54.5%, reactivity to non-structural NS3 antigen was noted in 27.3% (27/99) and to core antigen in 18.2% (18/99). No donors with isolated reactivity to NS4 were observed in our series.

Thrombosis in congenital deficiencies of AT III, protein C or protein S: a study of 44 children.

Congenital deficiency in coagulation inhibitors is a cause of hereditary thrombotic disease. The severity of symptoms is variable and depends on the type of deficit. In this paper, 44 children suffering from deep venous thrombosis, with a mean age of 5 years, were studied. A search for Lupus anticoagulant (LA) and coagulation inhibitor deficiency showed: 3/44 cases (6.8%) had protein S deficiency, 2/44 cases (4.5%) had protein C deficiency, 1/44 cases (2.3%) had deficiencies in both protein C and S; no cases of AT III deficiency and LA was positive in 2/44 cases (4.5%). Only 1 case of APC resistance out of 13 studied was found. Four family studies were performed and confirmed the congenital origin of the disorder.

[Deficiencies of physiological coagulation inhibitors in children with venous thromboses of the lower limbs. Prospective study of 6 cases]. / Déficts des inhibiteurs physiologiques de la coagulation dans les thromboses veineuses des membres inférieurs de l'enfant. Etude prospective de six cas.

Six patients aged 3 to 14 years with lower limb vein thrombosis were included in a prospective study of deficiencies in physiological coagulation inhibitors. The laboratory evaluation included standard hemostasis tests, tests for circulating anticoagulants, immunological and functional assays of protein C, protein S, and antithrombin III, and a study of fibrinolysis. A qualitative protein S deficiency with decreased fibrinolysis and protein C deficiency were found. The family study detected asymptomatic heterozygotes in both families investigated. No antithrombin III deficiency or circulating anticoagulants were found.

[Blood donors and anemia]. / Donneurs de sang et anémie.

Biological examinations of 2,630 blood donors showed that 8.6% of them were suffering with anemia. In 4% of the cases we discovered iron deficit and an hemoglobinopathy was found in 0.64% of all the cases.

[Galactosialidosis with Kayser-Fleischer's ring]. / La galactosialidose avec anneau de Kayser-Fleischer.

The case of a 4 year-old boy presenting with dysmorphic facies, hepatomegaly, splenomegaly, growth and psychomotor retardation is reported. Radiological pattern suggested a storage disease. Bone marrow differential cell count showed numerous storage cells with vacuolated lymphocytes. Enzymatic studies showed decreased beta-galactosidase and neuraminidase levels, leading to the diagnosis of galactosialidosis. This is the first Tunisian case reported, which differs from the other cases published by the presence of a Kayser-Fleischer ring.

[Genetic study of congenital afibrinogenemia. Review of 12 cases]. / Contribution à l'étude génétique de l'afibrinogénémie congénitale. A propos de douze cas.

Twelve cases of congenital afibrinogenemia in 11 families are reported. A family study was performed in six cases. The parents were genetically related in 8 of the 11 families. In half the cases another sibling had the disease. In every case the direct ascendants were unaffected. On the basis of results of plasma fibrinogen assays, "unprotected" heterozygotes with no more than 2.5 g/l fibrinogen and "protected" heterozygotes with normal fibrinogen levels were differentiated. Identification of "unprotected" heterozygotes is essential for genetic counselling. The reason for this variable phenotypic expression of congenital afibrinogenemia is unclear.

[A new case of relapsing hemolytic and uremic syndrome]. / A propos d'un nouveau cas de syndrome hémolytique et urémique a rechutes.

Relapsing hemolytic and uremic syndrome is infrequent in pediatric patients. The case of an infant who had HUS at 11 months of age and subsequently developed two relapses at 3 years and 3 1/2 years of age is reported. Outcome of the first episode was favorable, with no elements suggesting an increased risk for subsequent relapses. Conversely, the second and third episodes were followed by gradual deterioration of renal function and persistence of severe arterial hypertension. To clarify the epidemiology, clinical manifestations, histology, and outcome of HUS, the 24 pediatric cases of relapsing HUS reported in the English-language medical literature were reviewed. In this group, mean age at onset was 4 years 1 month (range 4 months-9 years) and number of relapses ranged from 1 to 14. Evidence of microangiopathy was the most common histologic finding (15/22) but non-specific lesions were seen in some instances. Opinions vary as to the prognosis of relapsing HUS; our case suggests that relapses are of adverse prognostic significance.

First case in Tunisia of PH1 positive chronic myelogenous leukemia in a 3 year old child.

The first case in Tunisia of a 3 year old child with adult type chronic myelogenous leukemia is presented. Diagnosis was based on clinical and biological presentation and detection of Philadelphia (PH1) chromosome. Chronic myelogenous leukemia in children represents 3.8% of all cases of chronic myelogenous leukemia detected in Tunisia between 1970 and 1990. Only one 3.5 year old case was reported without PH1 chromosome.