Abstract：
:Leishmaniasis, caused by protozoa belonging to the genus Leishmania, is an important public health problem found in >90 countries and with still limited options for treatment. Development of new anti-leishmanial drugs is an urgent need and the identification of new active compounds is a limiting factor that can be accelerated through large scale drug screening. This requires multiple steps and can be expensive and time consuming. Here, we propose an alternative approach for the colorimetric assessment of anti-Leishmania drug activity that can be easily scaled up. L. amazonensis and L. infantum cell lines were generated having the β-galactosidase (β-gal) gene integrated into their chromosomal 18S rRNA (ssu) locus. Both cell lines expressed high levels of β-gal and had their growth easily monitored and quantified colorimetrically. These two cell lines were then evaluated as tools to assess drug susceptibility and their use was validated through in vitro assays with Amphotericin B, which is routinely used against leishmaniasis. β-gal expression was also confirmed through flow-cytometry, another method of phenotypic detection. With these recombinant parasites, an alternative in vitro model of drug screening against cutaneous and visceral leishmaniasis is now available.
journal_name
J Microbiol Methodsjournal_title
Journal of microbiological methodsauthors
da Silva Santos AC,Moura DMN,Dos Santos TAR,de Melo Neto OP,Pereira VRAdoi
10.1016/j.mimet.2019.105732subject
Has Abstractpub_date
2019-11-01 00:00:00pages
105732eissn
0167-7012issn
1872-8359pii
S0167-7012(19)30303-3journal_volume
166pub_type
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