From: Leishmaniases diagnosis: an update on the use of immunological and molecular tools
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. |