**4. Conclusion**

more invasive samples such as lymph node aspirates [136]. Ferreira *et al*. (2012) used cPCR followed by kDNA-probe hybridization of blood samples, skin, marrow and conjunctival swabs. The use of swabs gave the best results for detection of infection in both symptomatic and asymptomatic dogs, showing to be a suitable sample for the molecular diagnosis of CVL. Furthermore, Di Muccio et al (2012) argue that the use of nested PCR from conjunctival swabs shown to be the least invasive procedure with the best performance for the diagnosis of CVL

Among other less invasive sample types investigated, Solano-Gallego et al (2007) evaluated urine samples with qPCR technique, but the results described showed positivity only in dogs with severe renal injury [160]. Naranjo et al. (2012) identified the presence of *Leishmania* sp. DNA in lacrimal glands of infected dogs using qPCR, with positive correlation between positivity and the presence of ocular lesions [161]. Recently, Belinchon-Lorenzo (2013), using qPCR, demonstrated the presence of *Leishmania* kDNA in the hair and keratinocytes of infected animals. According to the authors, the use of the non-invasive hair sample for the diagnosis of CVL would be advantageous because it is easy to collect, handle, transport, and store [135]. However, further studies should be conducted to determine the sensitivity of this method.

> Elleviti – Torino, Italy 26.80\* --- 63.00\* --- Scanelis - Toulouse, France --- --- 60.30\* ---

Toulouse, France --- --- --- 127.30\*

Laboratório Veterinário INNO – Braga, Portugal 20.60\* 54.40\* --- ---

Cornell University - Ithaca NY, USA 22.50 60.00 --- --- Hermes Pardini - Belo Horizonte MG, Brazil 17.20\* 60.20\* --- --- Análisis Biológicos– Chapecó SC, Brazil 9.40\* 42.15\* 72.25\* -- Laborlife - Rio de Janeiro RJ, Brazil 30.10\* 77.40\* --- ---

Centro de Investigación y Análisis Biológicos – Madrid, Spain 13.60\* 60.30\* 73.70\* ---

\*Prices in Euros (€) and Brazilian Real (R\$) converted in US dollars (U\$), quotation of November 14th of 2013. €1 = aprox.

**Table 3.** Cost of the main diagnostic test for CVL in different laboratories consulted in the second semester of 2013 in

**Tests Price Serology cPCR qPCR qPCR and**

28.00\* 41.20\* --- ---

19.20 --- --- ---

**Citology**

in asymptomatic animals [156].

258 Leishmaniasis - Trends in Epidemiology, Diagnosis and Treatment

**Laboratory – Country**

Laboratoire d'Anatomie Pathologique Vétérinaire du Sud-Ouest –

Instituto Nacional de Investigação Agrária e Veterinária, I.P. – Lisbõa, Portugal

Texas Veterinary Medical Diagnostic Laboratory – San Antonio TX, USA

U\$ 1.34 - €1 = aprox.R\$ 0.43

U\$

In summary, detailed clinical evaluation complemented with highly sensitive test allows proper identification of infected dogs in an endemic area. Evidence shows that the use of a rapid serological test associated with a molecular diagnostic test with high specificity, such as qPCR, is required for identification of all infected dogs, both asymptomatic and symptomatic. On the other hand, for sick dogs a correct diagnosis is necessary either to perform dog culling in countries where this measure is used as a control strategy of VL or to define treatment. In this case, a detailed clinical evaluation should be associated with biochemistry and hemato‐ logical tests to identify signs of renal and hepatic failure, in conjunction with a serological test to confirm animal clinical condition.
