Standardization of NASBA method by using 18s rRNA gene for Identification of Leishmania major parasite
Trauma Monthly: 14 (3); 137-142 Article Type: Research Article
S, Dalimi Asl
A, Foruzandeh Moghaddam
F. Standardization of NASBA method by using 18s rRNA gene for Identification of Leishmania major parasite,
Online ahead of Print
Aims: Leishmania major is a flagellated protozoan parasite, with more than 20 species and global distribution. This parasite can cause skin disease cutaneous leishmaniasis (CL) in humans. In diagnosis of the parasite, molecular methods are more sensitive and convenient than microscopic methods. Application of NASBA (Nucleic Acid Sequence-Based Amplification) method is shown to be high efficient for diagnosis of live parasite. In the present work, molecular isothermal method of NASBA was evaluated to identify live leishmania In vitro.
Materials & Methods: Parasites were cultured in RPMI and then their RNA was purified from promastigote stage. For evaluation of NASBA method in diagnosis of live parasite, the 18s rRNA gene of Leishmania major was amplified. The result band was investigated on agarose gel.
Results:Extracted RNA from parasite was 12.5kD and showed 3 bands of 24sα-, 24sβ rRNA and 18s rRNA. 18s rRNA gene of leishmania was appropriate for identification of Leishmania major in NASBA in the area of approximately 200bp.
Conclusion: The application of NASBA method is suitable for diagnosis of live Leishmania major. This method is relevant for evaluating effectiveness of course and treatment of anti leishmania drugs as well as early diagnosis of leishmania.
© 0, Author(s). This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.