title: Generation of stable L. major(+EGFP-LUC) and simultaneous comparison between EGFP and luciferase sensitivity.
abstract: Because of the lack of an accurate and sensitive tool to evaluate the parasitemia level, treatment or prevention of leishmaniasis remains an important challenge worldwide. To monitor and track leishmanial infection by two parameters in real time, we generated stably transgenic Leishmania that express a bi-reporter protein as fused EGFP and firefly luciferase. Using two reporter genes (egfp-luc) simultaneously increases the experimental sensitivity for detection/diagnosis, and in vitro quantification of parasites as well as real-time infection in mice. Through different specific tools, EGFP and LUC signals from the parasite were detectable and measurable within a mammalian host and promastigotes. Here, the LUC protein provided a higher level of sensitivity than did EGFP, so that infection was detectable at an earlier stage of the disease in the footpad (injection site) and lymph nodes by bioluminescence. These results depicted that: (1) both quantitative reporter genes, EGFP and LUC, could be simultaneously used to detect parasitemia in vitro and in vivo and (2) sensitivity of firefly luciferase was 10-fold higher than that of EGFP in promastigotes.
authors: Molecular Immunology and Vaccine Research Laboratory, Pasteur Institute of Iran, Tehran, Iran. Electronic address: Tahereh_t@yahoo.com.
authors: Tahereh
authors: T
authors: Taheri
authors: Molecular Immunology and Vaccine Research Laboratory, Pasteur Institute of Iran, Tehran, Iran; Fars Science and Research Branch, Islamic Azad University, Shiraz, Iran.
authors: Hana
authors: H
authors: Saberi Nik
authors: Molecular Immunology and Vaccine Research Laboratory, Pasteur Institute of Iran, Tehran, Iran.
authors: Negar
authors: N
authors: Seyed
authors: Molecular Immunology and Vaccine Research Laboratory, Pasteur Institute of Iran, Tehran, Iran.
authors: Fatemeh
authors: F
authors: Doustdari
authors: Department of Virology, Pasteur Institute of Iran, Tehran, Iran.
authors: Mohammad-Hossein
authors: MH
authors: Etemadzadeh
authors: Department of Medical Biotechnology, Pasteur Institute of Iran, Tehran, Iran.
authors: Fatemeh
authors: F
authors: Torkashvand
authors: Molecular Immunology and Vaccine Research Laboratory, Pasteur Institute of Iran, Tehran, Iran.
authors: Sima
authors: S
authors: Rafati
mesh_terms: D000666:Amphotericin B / Q000494:pharmacology / Q000627:therapeutic use; D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology / Q000627:therapeutic use; D005260:Female; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D018320:Leishmania major / Q000187:drug effects / Q000235:genetics*; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis / Q000188:drug therapy / Q000469:parasitology*; D049409:Luciferases, Firefly / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D030781:Organisms, Genetically Modified; D013997:Time Factors; D014162:Transfection
chemical_list: D000981:Antiprotozoal Agents; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D000666:Amphotericin B; D049409:Luciferases, Firefly
keywords: Amphotericin B; Green fluorescent protein; Leishmania major; Leishmaniasis; Luciferase