Tags

Type your tag names separated by a space and hit enter

Novel approach to in vitro drug susceptibility assessment of clinical strains of Leishmania spp.
J Clin Microbiol. 2012 Jul; 50(7):2207-11.JC

Abstract

Resistance to antimonial drugs has been documented in Leishmania isolates transmitted in South America, Europe, and Asia. The frequency and distribution of resistance to these and other antileishmanial drugs are unknown. Technical constraints have limited the assessment of drug susceptibility of clinical strains of Leishmania. Susceptibility of experimentally selected lines and 130 clinical strains of Leishmania panamensis, L. braziliensis, and L. guyanensis to meglumine antimoniate and miltefosine was determined on the basis of parasite burden and percentage of infected U-937 human macrophages. Reductions of infection at single predefined concentrations of meglumine antimoniate and miltefosine and 50% effective doses (ED(50)s) were measured and correlated. The effects of 34°C and 37°C incubation temperatures and different parasite-to-host cell ratios on drug susceptibility were evaluated at 5, 10, and 20 parasites/cell. Reduction of the intracellular burden of Leishmania amastigotes in U-937 cells exposed to the predefined concentrations of meglumine antimoniate or miltefosine discriminated sensitive and experimentally derived resistant Leishmania populations and was significantly correlated with ED(50) values of clinical strains (for meglumine antimoniate, ρ = -0.926 and P < 0.001; for miltefosine, ρ = -0.906 and P < 0.001). Incubation at 37°C significantly inhibited parasite growth compared to that at 34°C in the absence of antileishmanial drugs and resulted in a significantly lower ED(50) in the presence of drugs. Susceptibility assessment was not altered by the parasite-to-cell ratio over the range evaluated. In conclusion, measurement of the reduction of parasite burden at a single predetermined drug concentration under standardized conditions provides an efficient and reliable strategy for susceptibility evaluation and monitoring of clinical strains of Leishmania.

Authors+Show Affiliations

Centro Internacional de Entrenamiento e Investigaciones Médica, Cali, Colombia.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Evaluation Study
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

22518860

Citation

Fernández, Olga, et al. "Novel Approach to in Vitro Drug Susceptibility Assessment of Clinical Strains of Leishmania Spp." Journal of Clinical Microbiology, vol. 50, no. 7, 2012, pp. 2207-11.
Fernández O, Diaz-Toro Y, Valderrama L, et al. Novel approach to in vitro drug susceptibility assessment of clinical strains of Leishmania spp. J Clin Microbiol. 2012;50(7):2207-11.
Fernández, O., Diaz-Toro, Y., Valderrama, L., Ovalle, C., Valderrama, M., Castillo, H., Perez, M., & Saravia, N. G. (2012). Novel approach to in vitro drug susceptibility assessment of clinical strains of Leishmania spp. Journal of Clinical Microbiology, 50(7), 2207-11. https://doi.org/10.1128/JCM.00216-12
Fernández O, et al. Novel Approach to in Vitro Drug Susceptibility Assessment of Clinical Strains of Leishmania Spp. J Clin Microbiol. 2012;50(7):2207-11. PubMed PMID: 22518860.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Novel approach to in vitro drug susceptibility assessment of clinical strains of Leishmania spp. AU - Fernández,Olga, AU - Diaz-Toro,Yira, AU - Valderrama,Liliana, AU - Ovalle,Clemencia, AU - Valderrama,Mabel, AU - Castillo,Harry, AU - Perez,Mauricio, AU - Saravia,Nancy Gore, Y1 - 2012/04/18/ PY - 2012/4/21/entrez PY - 2012/4/21/pubmed PY - 2012/10/18/medline SP - 2207 EP - 11 JF - Journal of clinical microbiology JO - J Clin Microbiol VL - 50 IS - 7 N2 - Resistance to antimonial drugs has been documented in Leishmania isolates transmitted in South America, Europe, and Asia. The frequency and distribution of resistance to these and other antileishmanial drugs are unknown. Technical constraints have limited the assessment of drug susceptibility of clinical strains of Leishmania. Susceptibility of experimentally selected lines and 130 clinical strains of Leishmania panamensis, L. braziliensis, and L. guyanensis to meglumine antimoniate and miltefosine was determined on the basis of parasite burden and percentage of infected U-937 human macrophages. Reductions of infection at single predefined concentrations of meglumine antimoniate and miltefosine and 50% effective doses (ED(50)s) were measured and correlated. The effects of 34°C and 37°C incubation temperatures and different parasite-to-host cell ratios on drug susceptibility were evaluated at 5, 10, and 20 parasites/cell. Reduction of the intracellular burden of Leishmania amastigotes in U-937 cells exposed to the predefined concentrations of meglumine antimoniate or miltefosine discriminated sensitive and experimentally derived resistant Leishmania populations and was significantly correlated with ED(50) values of clinical strains (for meglumine antimoniate, ρ = -0.926 and P < 0.001; for miltefosine, ρ = -0.906 and P < 0.001). Incubation at 37°C significantly inhibited parasite growth compared to that at 34°C in the absence of antileishmanial drugs and resulted in a significantly lower ED(50) in the presence of drugs. Susceptibility assessment was not altered by the parasite-to-cell ratio over the range evaluated. In conclusion, measurement of the reduction of parasite burden at a single predetermined drug concentration under standardized conditions provides an efficient and reliable strategy for susceptibility evaluation and monitoring of clinical strains of Leishmania. SN - 1098-660X UR - https://www.unboundmedicine.com/medline/citation/22518860/Novel_approach_to_in_vitro_drug_susceptibility_assessment_of_clinical_strains_of_Leishmania_spp_ L2 - http://jcm.asm.org/cgi/pmidlookup?view=long&amp;pmid=22518860 DB - PRIME DP - Unbound Medicine ER -