A rapid high-resolution melting method for differentiation of Leishmania species targeting lack gene.

Objectives: The aim of this research is to verify that if lack gene can be used for differentiation of Leishmania under HRM assay.

Methods: Two specific primers were designed targeting polymorphic sites on the lack gene sequence. DNA from promastigotes of six species of Leishmania based on reference strains were tested following a HRM protocol. We also tested ten Chinese isolates in blind to validate our method.

Results: Combined with amplicon of the two primers, the six reference strains can be easily discriminated without the effect of initial concentration of DNA templates. Ten Chinese isolates detected by our HRM method resulted in full accord with the standard identification results in previous study.

Conclusion: HRM is a rapid and reproducible method to discriminate different Leishmania species and lack gene is a potential novel biological characteristic for easy differentiation of Leishmania isolates in China.