**3.4 Molecular biology**

Up to 2008, our laboratory was not equipped for molecular biology techniques. In 2007, as part of a collaborative project on "molecular tools for accurate diagnosis and better assessment of leishmaniases", codirected by the LEEP and our own laboratory and financed by the IAEA, a molecular biology unit was set up in our department and has just started working. In the meanwhile we have been collaborating with the molecular biology unit of the LEEP, mainly for identifying *Leishmania* strains. The first study, conducted in 2005, aimed at identifying strains isolated from patients suspected for having SCL, by using K-DNA-PCR according to Smyth et al., 1992 and PCR-RFLP according to Guerbouj et al., 2001 (Ben Said et al., 2006).

Retrospective Analysis of Leishmaniasis in

**0**

Sidi Bouzid

Mahdia

Kairouan

Sousse

Others : Monastir, Gafsa, Kasserine, Sfax, Tataouine, Tozeur, Gabes, Kébili

governorate and delegation.

**50**

**100**

**150**

**200**

**250**

**Number of cases** 

**300**

**350**

**400**

**450**

Central Tunisia: An Update on Emerging Epidemiological Trends 239

**1986 1988 1990 1992 1994 1996 1998 2000 2002 2004 2006 2008 2010**

Governorate Delegation Nb/delegation Nb (% /governorate)

241 33 32


610 (32.6 %)

494 (26.4 %)

369 (19.7 %)

306 (16.3 %)

Fig. 9. Annual distribution of 2051 zoonotic cutaneous leishmaniasis cases (1986-2010).

Ouled Haffouz Jelma Sidi Bouzid city Others

Chorbène Souassi Hbira Ouled Chamekh Others

Nasrallah Bouhajla Kairouan south Hadjeb layoun Others

> Sidi El Heni Msaken Others

Table 3. Distribution of 1873 zoonotic cutaneous leishmaniasis cases according to

TOTAL - - 1873

Later, additional strains isolated from CL patients were typed by a novel Multiplex PCR (Saadi et al., in preparation) as part of the IAEA project that is still ongoing.

In addition, specimens from patients with lesions highly evocative of CL but found negative in parasitological examination were addressed for PCR to the laboratory of parasitology of the Faculty of Pharmacy, Monastir, where a molecular biology unit has just been set up. PCR was performed according to Chargui et al., 2005.
