Ar Niazi, F Koohsar, F Ghaffarifar, H Ziaei-Hezarjaribi,, F Mesgarian, on Jorjani,
Volume 8, Issue 2 (summer 2014[PERSIAN] 2014)
Abstract
Abstract
Background and Objective: Culture, microscopic method is a gold standard method for identification of Lishmania parasite. The use of Molecular methods such as RT- PCR compared to microscopic methods has a higher sensitivity and specificity however, it is not widely used due to its expensive equipment and the time requested. The use of nucleic acid sequence based amplification (NASBA) method is highly valuable for diagnosis of live parasite because there is no need for to use Thermo cycler. We aimed to assess sensitivity and specificity of NASBA for molecular detection of cutaneous Leishmaniasis.
Material and Methods: First, the RNA was extracted from 28 skin biopsies suspected cutaneous Leishmaniasis. Then, by means of specific primers designed for 18srRNA region, this region was amplified using NASBA isothemal amplification. To increase the sensitivity, the product was electroforesed in TBE (IX) buffer, using Syber Gold Flourecent probes. Using specific primers, RT- PCR was conducted on the samples too.
Result: For diagnosis of Leishmania parasites, NASBA and RT-PCR had the sensitivity of 81% and 51%, respectively, and specificity of 100%.
Conclusion: NASBA isothermal method with high sensitivity and specificity can be applied for identification of cutaneous leishmaniasis.
Keywords: Cutaneous Leishmanisis, NASBA, 18S rRNA
Mojtaba Raeisi, Kamal Mirkarimi, Behrooz Jannat, Bahman Rahimi Esboei, Abdol Sattar Pagheh, Zahra Mehrbakhsh, Fatemeh Ghaffarifar, Oghlniaz Jorjani, Masoud Foroutan,
Volume 14, Issue 4 (Jul-Aug 2020)
Abstract
Background and objectives: Leishmaniasis is a tropical disease caused by protozoan parasites from the genus Leishmania. In this study, we aimed at investigating the in vitro anti-leishmanial effect of essential oils of Rosmarinus officinalis, Mentha pulegium, Foeniculum vulgare, Lippia citriodora and Pelargonium graveolens.
Methods: The essential oils were prepared from freshly dried and powdered plants with steam-distilled water. Iranian strain of Leishmania promastigotes was cultured in RPMI medium and the inhibitory effects of different concentrations (25, 32, 62.5, 125, 250, 500 and 1000 μg/ml) of the essential oils were investigated at 24, 48 and 72 hours. The number of live parasites before and after treatment with the essential oils was counted by trypan blue 10% staining and using neobar lam.
Results: The essential oils significantly decreased the number of promastigotes in a dose-dependent manner (P<0.05). However, the inhibitory effects of F. vulgare and R. officinalis essential oils were more profound compared to other essential oils. Moreover, concentrations of 500 and 1000 μg/ml of these two essential oils exerted equal and more anti-leishmanial potency compared to glucantime, the first-line drug used for treatment of leishmaniasis.
Conclusion: Based on the results, it is recommended to evaluate the in vivo anti-leishmanial effects of the tested essential oils, particularly F. vulgare and R. officinalis.
