**3.2.1 Parasitological diagnosis**

236 Current Topics in Tropical Medicine

2004; Pousse et al. 1995). In HIV Tunisian patients, VL is infrequent; it is best diagnosed by

Fig. 8. Actual distribution of visceral leishmaniasis in Tunisia.

The laboratory of parasitology of Sousse, the third major Tunisian city which is located in the central, coastal part of Tunisia, was first created in 1986. It can be considered as a central laboratory ensuring the diagnosis of parasitic diseases' cases including leishmaniasis, sent by generalists, dermatologists, pediatricians or hematologists either from the same hospital or other hospitals mainly from governorates of central Tunisia like Kairouan, Sidi Bouzid and Mahdia. Results are usually sent back to the referring doctors so the patients can be managed as appropriate. Since its creation, the laboratory has been involved in the

For purposes of this retrospective study, VL and CL patients' data, including age, sex, geographical origin and the likely place of contamination, clinical presentation and

The diagnosis of CL is usually achieved by the demonstration of amastigotes in Giemsastained smears from the fluid obtained by scraping the edges of the cutaneous lesion with a

**3. Materials and methods** 

diagnosis of all forms of leishmaniasis.

evolution without or after treatment were collected.

**3.1 Diagnosis of cutaneous leishmaniasis** 

PCR (Kallel et al., 2007).

Diagnosis of VL is usually based on the demonstration of amastigotes in Giemsa stained bone-marrow aspirates. However, in our laboratory, bone-marrow smears are mainly carried out and read in doubtful cases, because they usually are performed and interpreted by the haematologists and sometimes the pediatricians themselves and the results of the examination are later sent to us together with sera specimens.

For the purpose of parasitological diagnosis of VL, since 1996, we have been using the cytoconcentration of peripheral blood according to the technique of Petithory et al, 1997. Cytoconcentration is usually practiced first, and when positive the painful bone marrow aspiration is no more needed and the diagnosis of VL is established.
