[Contribution of Real-Time PCR in the Diagnosis of Cutaneous Leishmaniasis: Experience of the Pasteur Institute of Tunis]. / Contribution de la PCR en temps réel dans le diagnostic biologique de la leishmaniose cutanée : expérience de l'institut Pasteur de Tunis.

The aim of this study was to assess the performance of real-time PCR (qPCR) in the diagnosis of cutaneous leishmaniasis (CL). Culture, direct microscopic examination (DE) and qPCR were performed on dermal exudate samples collected from 235 confirmed CL cases. The qPCR was found to be more sensitive than other diagnostic techniques and was able to correct the diagnosis in 49 patients (20.9%) with negative dermal smears. Median parasitic load (PL) of the 49 dermal exudates with negative DE was lower than that of positive ones in microscopy. This suggests that PL likely impact the sensitivity of microscopy. On the other hand, qPCR was performed on DNA extracts of scraped products collected from the 23 out of 49 archived negative Giemsa-stained slides and showed 11 positive. Parasitic loads in the latter smears were lower than those in corresponding exudates. The results highlight qPCR relevance for the diagnosis of CL and recommend its use directly on dermal exudates collected from CL lesions.

L'objectif de ce travail était d'évaluer la place de la PCR en temps réel (qPCR) dans le diagnostic de la leishmaniose cutanée (LC). Des sucs dermiques prélevés chez 235 cas confirmés de LC ont été traités par examen microscopique direct (ED), culture et qPCR, et les résultats analysés. La qPCR s'est révélée plus sensible que les autres techniques diagnostiques permettant de redresser le diagnostic chez 49 patients (20,9 %). Les charges parasitaires (CP) des 49 sucs dermiques correspondants étaient inférieures à celles de prélèvements microscopiquement positifs suggérant que la CP serait un facteur limitant la sensibilité de l'ED. La qPCR a été également pratiquée sur les produits de grattage de 23 des 49 frottis négatifs. Elle a permis d'en détecter 11 positifs avec des CP inférieures à celles dans les sucs dermiques correspondants. Ces résultats recommandent l'utilisation de la qPCR pour le diagnostic de la LC et privilégient sa réalisation directement sur le suc dermique.

[Management of Toxoplasmic Seroconversion in the Third Trimester of Pregnancy in Tunisia]. / Particularités de la prise en charge en Tunisie des séroconversions toxoplasmiques du troisième trimestre de grossesse.

The purpose of our study was to report the particularities of the management of toxoplasmosis seroconversion cases occurred during the third trimester of pregnancy and diagnosed in the Institute Pasteur of Tunis. The study was conducted in the Laboratory of Parasitology-Mycology of the Institute Pasteur of Tunis between January 2005 and December 2017. A total of 27 cases of toxoplasmosis seroconversion during the third trimester were included. Prenatal diagnosis was performed in five cases. PCR was positive in one case. Pyrimethamine-sulfadoxine was prescribed in one case with positive PCR and in another case as soon as maternal infection was confirmed. Spiramycine was prescribed in 24 cases for the duration of the pregnancy. One woman did not take any treatment because seroconversion was diagnosed just before delivery. Twenty newborns had congenital toxoplasmosis: 19 cases were diagnosed by serology and one case was diagnosed after amniocentesis. Two newborns (10%) were symptomatic at birth. All the newborns had neither clinical nor radiological signs during the follow-up.

L'objectif de notre travail était de rapporter les particularités de la prise en charge d'une série de cas de séroconversion toxoplasmique du troisième trimestre de grossesse diagnostiquée à l'institut Pasteur de Tunis. Il s'agit d'une étude rétrospective colligée au laboratoire de parasitologie-mycologie de l'institut Pasteur de Tunis entre janvier 2005 et décembre 2017, incluant 27 femmes enceintes ayant présenté une séroconversion toxoplasmique au cours du troisième trimestre de la grossesse diagnostiquée au cours de leurs suivis sérologiques. Cinq femmes ont bénéficié d'un diagnostic anténatal. La PCR (polymerase chain reaction) s'est révélée positive dans un seul cas. Deux femmes ont été mises sous pyriméthamine-sulfadoxine dont une avait une PCR positive. La spiramycine a été instituée chez 24 femmes jusqu'à l'accouchement. Une femme n'a reçu aucun traitement, la séroconversion ayant été diagnostiquée la veille de l'accouchement. Le diagnostic de la toxoplasmose congénitale a été retenu chez 20 nouveau-nés (74 %) dont un en anténatal et 19 à la naissance. Deux étaient symptomatiques (10 %) à la naissance. Aucune manifestation clinique ou radiologique n'a été observée au cours de leurs suivis.

[Toxoplasmic infections in pregnancy: about 94 cases diagnosed at the Pasteur Institute of Tunis]. / Infections toxoplasmiques au cours de la grossesse: à propos de 94 cas diagnostiqués à l'institut Pasteur de Tunis.

OBJECTIVES: The objectives of this study were to estimate the risk of maternal-fetal transmission of toxoplasmosis and its consequences on the fetus and to emphasize the importance of follow-up of newborns in Tunisia. PATIENTS AND METHODS: It was a retrospective study of 94 cases of pergravidic toxoplasmic seroconversion who were diagnosed and followed in the Laboratory of Parasitology of Pasteur Institute of Tunis between 2005 and 2010. RESULTS: In our series, amniocentesis was performed for 60 parturients. Among the amniotic fluid tested, research of toxoplasmosis DNA by PCR was positive in 12 cases (12/60, 20 %). Twenty-six cases of congenital toxoplasmosis were diagnosed with 14 postnatal cases. The rate of maternal-fetal transmission of Toxoplasma gondii was 27.6 % (26/94). This risk increases with gestational age, from 19 % at seroconversion of the 1st quarter to 29.4 % in the 2nd quarter and 44.4 % in the 3rd trimester. Monitoring of newborns with congenital toxoplasmosis showed that only 3 children were symptomatic. There were 2 cases of toxoplasmic chorioretinitis and a case of brain damage. Under serological monitoring of newborns, 21 cases were lost to follow-up and monitoring was stopped for 29 after decrease of anti-toxoplasmic IgG. DISCUSSION AND CONCLUSION: The prenatal diagnosis allowed to decrease the severe forms of congenital toxoplasmosis in Tunisia. Nevertheless, it is always necessary to raise the problem of the significant number of newborn children whose follow-up is incomplete.

[Toxoplasmosis mother-to-child screening: study of cases followed in the Pasteur Institute of Tunis (2007-2010)]. / Dépistage de la toxoplasmose materno-foetale : étude des cas suivis à l'Institut Pasteur de Tunis (2007-2010).

Toxoplasmosis when occurring during pregnancy can be transmitted to the fetus and lead to congenital toxoplasmosis (CT). Therefore, pregnant women are a risk group, for which it is necessary to determine the serologic profile. The objective of this study is to determine the serologic profile of toxoplasmosis in pregnant women followed at the Parasitology Laboratory of the Pasteur Institute in Tunis, to establish the prevalence of toxoplasmic infections during pregnancy and the incidence of the CT, noting the difficulties faced in the interpretation of serological results. This is a retrospective study concerning 2833 toxoplasmic serologies practiced on 2070 pregnant women, followed at the Parasitology-Mycology Laboratory of the Pasteur Institute of Tunis, between 2007 and 2010. Serological diagnosis of toxoplasmosis was done by ELISA (Enzyme Linked Immunosorbent Assay) for the detection of Immunoglobulin (Ig) G and M and the study of toxoplasmosis IgG avidity. Prenatal diagnosis was performed for 58 women by amniotic fluid sampling. Toxoplasma gondii was detected by Polymerase Chain Reaction (PCR). At birth, the diagnosis of congenital toxoplasmosis was established based on serology. The toxoplasmic serologies carried out have shown that 45.6% of the pregnant women were formerly immunized while 49.6% had a negative serology. A toxoplasmosis primary infection acquired during pregnancy was detected in 79 cases (3.8%). Among them, 33% had a true seroconversion while 67% had a recent toxoplasmosis infection in view of the positivity of IgG and IgM on the first sample with a low index of avidity (IA). For 21 parturients whose serology showed the presence of IgG, IgM and an intermediate or high IA. Among the 58 parturients in whom prenatal diagnosis was performed, PCR was positive in four cases. After birth, six cases of congenital toxoplasmosis were detected by serology.

[Ileocecal ameboma: a case diagnosed by pathology and molecular biology]. / Amoebome iléocaecal: un cas diagnostique par l'anatomie pathologique et la biologie moléculaire.

Colonic ameboma is a rare benign inflammatory tumor due to the infection by Entamoeba histolytica and poses frequently the problem of colon cancer. We report a case of a 52 year-old patient who presented a cecal amoeboma revealed by a painful mass in the right iliac fossa. Radiologic and endoscopic examinations depicted a parietal thickening of the right colon and the cecum. A presumptive diagnosis of colon cancer was firstly discussed. Confirmation of ameboma was made on pathological examination, PCR and serology.

[Congenital toxoplasmosis following infection occurring late in pregnancy]. / Toxoplasmose congénitale faisant suite à une primo-infection maternelle en fin de grossesse.

Monthly serological screening of non immune pregnant women is recommended for prevention of congenital toxoplasmosis. However, this screening is often interrupted before delivery. We report a case of congenital toxoplasmosis following infection occurring late in pregnancy. This documented case highlights the need for a final routine serological test, 2-3 weeks post-partum for all seronegative pregnant women. In fact, the screening of congenital toxoplasmosis cases allows the early administration of specific treatment that avoids later severe complications such as chorioretinitis.

Case of fatal congenital toxoplasmosis associated with I/III recombinant genotype.

We report a case of fatal congenital toxoplasmosis case in Tunis (North of Tunisia) associated with I/III recombinant genotype. The Toxoplasma gondii strain was isolated from placenta and characterized molecularly by a multilocus typing (3'SAG2, 5'SAG2, SAG3, AK69, APICO, and UPRT1) and a sulfadiazine resistance mutation analysis. The isolate was shown to be a wild drug sensitive I/III recombinant strain.

[Congenital toxoplasmosis: clinical and biological analysis of 11 cases in Tunisia]. / La toxoplasmose congénitale en Tunisie: analyse clinique et biologique de 11 cas.

Early diagnosis of congenital toxoplasmosis (CT) is necessary to prevent serious complications. The CT is diagnosed by the identification of the parasite in amniotic fluid during pregnancy or at birth by detection of antibodies synthesized by the fetus. The aim of this study was to determine the clinical and biological particularities of CT in a cohort of 11 cases observed in Tunisia and to evaluate the performance of the prenatal and neonatal diagnostic techniques that were used. In all patients, the presumed date of maternal contamination was determined based on the mother's serological data. Neonatal serological screening included assays for immunoglobulins (Ig) G and IgM by enzyme-linked-immuno-sorbent assay (ELISA), IgM by immuno-sorbent-agglutination-assay (ISAGA), and comparison of the mother-baby immunological profile by western blot IgG and IgM. Seven out of the 11 cases had been diagnosed prenatally; only 3 (43%) had a positive polymerase chain reaction (PCR) test. Ten newborns out of 11 had serological criteria of fetal infection; western blot results were positive in all of them, ISAGA was positive in 2 and ELISA in none. The newborn who had a negative serology had been treated in utero. The date of positivity of the western blot test varied: 6 cases at birth, 2 at 12 days of life and 2 at 1 month of life. The 2 patients who had positive ISAGA results had been contaminated during the 3rd trimester of gestation.