Here, we determined the Toxoplasma gondii genotype in amniotic fluid, placenta, and cerebrospinal fluid samples from 14 congenital toxoplasmosis cases in Tunisia, North Africa. Direct genotypic characterization of T. gondii strains was performed by polymerase chain reaction (PCR) amplification of six genetic markers (3'SAG2, 5' SAG2, SAG3, BTUB, GRA6, and APICO) and thereafter, was analyzed by restriction fragment-length polymorphism (RFLP). Samples were sequenced to resolve strain type whenever there were unclear enzyme digestion results. Multilocus analysis revealed that only one specimen harbored the type I allele in all studied loci, whereas the 13 others gave mixed genotype results with different alleles at different markers. Seven specimens produced RFLP profile of the recombinant strains I/III, and three produced a profile of I/II recombinant strains. The last three specimens produced complex digestion patterns. In these cases, sequence analysis revealed double peaks at known polymorphic sites, indicating the presence of multiple alleles.

BackgroundClinical manifestation due to infection by Toxoplasma gondii is closely linked to the infecting strain of the parasite. Several genetic markers are available to determinate its genotype but few of them are able to discriminate between the three predominant lineages, namely types I, II and III. The number of markers decreases when atypical, recombinant/mixed genotypes need to be identified.FindingsIn our study, the contribution of sequence polymorphisms in the AK69 gene as typing markers for T. gondii was investigated for the first time in an epidemiological study. The coding region of the marker was amplified, sequenced and aligned for different Toxoplasma strains. The identified nucleotide polymorphism at 12 positions was able to highly discriminate between the different congenital toxoplasmosis Tunisian strains. Moreover the high detection sensitivity level of the marker enabled unambiguous identification of mixed/recombinant genotypes directly.ConclusionIt can be, thus, very useful for direct typing in areas where such genotypes are frequently encountered, mainly in the African continent.

We report here a case of simultaneous cutaneous and visceral manifestations due to Leishmania L. infantum diagnosed in an immunocompetent adult. We describe a 74-year-old woman from Tunis, Tunisia, who presented a biologically confirmed visceral leishmaniasis infection concomitant with arm ulceration which appeared 2 years before. Leishmania DNA was detected by ITS PCR in both buffy coat and dermal scrapping of the arm lesion. Sequencing revealed that the 2 isolated strains corresponded to L. infantum and were 100% identical. The symptoms of visceral leishmaniasis responded to amphotericin B with rapid healing. However, the skin lesion did not improve although Leishmania PCR on dermal sample became negative. This location is probably secondarily to lymphatic or blood dissemination during the systemic visceral leishmaniasis infection. It would be favored by the inflammatory environment induced by the basal cell carcinoma subsequently diagnosed.

Background An immunochromatography technology (ICT) rapid diagnostic test, the Toxoplasma ICT IgG‐IgM®, was recently developed. Our aim was to study its contribution to establish accurately the Toxoplasma immune status in Tunisian pregnant women using Western blot (WB) Toxo II IgG® as a reference technique. Methods Thirty‐nine sera were selected for the study from among 2,615 which were already tested by IgG and IgM ELISA. They displayed equivocal IgG titres (4.4–9 IU/ml) in absence of IgM (19 sera) or IgM anti‐Toxoplasma antibodies in absence of IgG (titre <4.4 IU/ml) (20 sera). All these sera were additionally tested by WB Toxo II IgG®. Results Immunochromatography technology Sensitivity in the detection either of low IgG titres in absence of IgM or of specific anti‐Toxoplasma IgM was 100%. Only one serum with equivocal IgG titre by ELISA and negative with Toxo II IgG® test revealed positive in ICT. However, this serum showed a P30 band in WB analysis. On the other hand, three sera positive in ELISA IgM and negative in ELISA IgG revealed positive in ICT and negative in WB Toxo II IgG®, the reference test. Conclusion Results confirm the high sensitivity of Toxoplasma ICT IgG‐IgM® in detecting both specific anti‐Toxoplasma IgG and IgM, and highlight the usefulness of this rapid test as a first or second‐line Toxoplasma serological test in pregnant women.