title: Low-cost liquid medium for in vitro cultivation of Leishmania parasites in low-income countries.
abstract: BACKGROUND
  Prompt laboratory diagnosis and initiation of treatment are effective components of leishmaniasis control. Detection of Leishmania parasites by ex-vivo culture of lesion scrapings is considered a definitive diagnostic method preceding initiation of treatment.
  
  
  OBJECTIVE
  A pilot study to find alternative medium that could reduce the cost of culturing from patient lesions for diagnosing leishmaniasis.
  
  
  METHODS
  GALF-1 medium was formulated in our lab from locally available inexpensive solutions and powders in the presence of urine from healthy individuals. Amastigote to promastigote transformation, recovery of parasites after cryopreservation, cost and mass cultivation was compared using the following media: GALF-1, RPMI 1640, and conventional Locke's semi-solid medium (LSSM), a modifications of Novy-MacNeal-Nicolle culture media, which uses Locke's solution as an overlay
  
  
  RESULTS
  GALF-1 preparation was cheap and the components available in low-income countries such as Ethiopia. Preparation was simple, not requiring autoclaving and extra distilled water. GALF-1 was able to transform amastigotes from Ethiopian patients' samples and could be used to cultivate promastigotes in large quantities. GALF-1 decreased Leishmania culture costs by approximately 80-95% compared to LSSM and RPMI 1640, respectively. Promastigotes cultured with GALF-1 could be cryopreserved in liquid nitrogen with comparable re-culture potential.
  
  
  CONCLUSIONS
  Affordability of diagnostic assays is a key issue for endemic resource-poor countries and the possibility to cut the cost of the efficient culture method for diagnosis through the use of inexpensive, locally formulated reagents could improve the diagnosis of leishmaniasis in Ethiopia and in other low-income countries.
authors.affiliation: Department of Infectious and Non-infectious Diseases Research, Ethiopian Health and Nutrition Research Institute (EHNRI), Addis Ababa, Ethiopia.
authors.forename: Geremew
authors.initials: G
authors.lastname: Tasew
authors.forename: Amha
authors.initials: A
authors.lastname: Kebede
authors.forename: Dawit
authors.initials: D
authors.lastname: Wolday
authors.forename: Endalamaw
authors.initials: E
authors.lastname: Gadisa
authors.forename: Sven
authors.initials: S
authors.lastname: Britton
authors.forename: Liv
authors.initials: L
authors.lastname: Eidsmo
authors.forename: Hannah
authors.initials: H
authors.lastname: Akuffo