Table 2.

Advantages and limitations of molecular methods used in leishmaniases diagnosis

Method	Advantage	Limitation
Conventional PCR (cPCR)	High sensitivity, specificity and accurate results. Many applications in molecular analysis. Easy diagnostic interpretation.	Unable to quantify the target DNA. Qualitative test. Time consuming. Limited detection range of some assays.
Quantitative real-time PCR (qPCR)	Higher sensitivity, specificity and security, quantitative capacity and speedy results. Possibility of species differentiation by melting temperature.	High cost due to equipment (thermocycler). Difficulty in interpreting the results, needing thus of a well-trained operator.
Nested-PCR (nPCR)	Higher specificity and sensitivity. Useful technique for studying the molecular epidemiology in the field.	Time consuming and higher cost. Unable to quantify the target DNA. Qualitative test.
Quantitative Nucleic Acid Sequence-Based Assay (QT-NASBA)	High specificity. It is based on an isothermal reaction and thus overcomes the need for a thermocycler; Ideal for lower-tech laboratories. Quantitative capacity. Indicated to detect active diseases; RNA detection.	It uses electrochemiluminescence as tool of detection, which involves more handling steps and procedure time. Assays developed only for RNA detection. . Few studies yet.
NASBA coupled with oligochromatography (NASBA-OC)	High specificity. Speedy results. There is no need of complex laboratorial structure. Simple dipstick format for the detection of amplification products. RNA detection.	Unable to quantify the target RNA. Assays developed only for RNA detection. Few studies yet.
Loop-Mediated Isothermal Amplification (LAMP)	High sensitivity. Low cost. Isothermal reaction, there is no need for a thermocycler. The temperature stability of the reagents enables its use in field conditions.	Unable to quantify the target DNA. Qualitative test. Few studies yet.