Low-cost liquid medium for in vitro cultivation of Leishmania parasites in low-income countries.
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.
Tasew
Geremew
G
Department of Infectious and Non-infectious Diseases Research, Ethiopian Health and Nutrition Research Institute (EHNRI), Addis Ababa, Ethiopia.
Kebede
Amha
A
Wolday
Dawit
D
Gadisa
Endalamaw
E
Britton
Sven
S
Eidsmo
Liv
L
Akuffo
Hannah
H