FN Archimer Export Format
PT J
TI Screening and Characterization of RAPD Markers in Viscerotropic Leishmania Parasites
BT 
AF MKADA-DRISS, Imen
   LAHMADI, Ramzi
   CHAKROUN, Ahmed S.
   TALBI, Chiraz
   GUERBOUJ, Souheila
   DRISS, Mehdi
   ELAMINE, Elwaleed M.
   CUPOLILLO, Elisa
   MUKHTAR, Moawia M.
   GUIZANI, Ikram
AS 1:1;2:1;3:1;4:1;5:1;6:1;7:2;8:3;9:2;10:1;
FF 1:;2:;3:;4:;5:;6:;7:;8:;9:;10:;
C1 Univ Tunis el Manar, Lab Epidemiol Mol & Pathol Expt LR11IPT04, Lab Epidemiol & Ecol Parasitaire LR00SP04, Inst Pasteur Tunis, Tunis, Tunisia.
   Univ Khartoum, Inst Endem Dis, Khartoum, Sudan.
   Inst Oswaldo Cruz, Lab Pesquisas Leishmaniose, BR-20001 Rio De Janeiro, Brazil.
C2 UNIV TUNIS, TUNISIA
   UNIV KHARTOUM, SUDAN
   INST OSWALDO CRUZ, BRAZIL
IN DOAJ
IF 3.234
TC 4
UR https://archimer.ifremer.fr/doc/00274/38559/37081.pdf
LA English
DT Article
AB Visceral leishmaniasis (VL) is mainly due to the Leishmania donovani complex. VL is endemic in many countries worldwide including East Africa and the Mediterranean region where the epidemiology is complex. Taxonomy of these pathogens is under controversy but there is a correlation between their genetic diversity and geographical origin. With steady increase in genome knowledge, RAPD is still a useful approach to identify and characterize novel DNA markers. Our aim was to identify and characterize polymorphic DNA markers in VL Leishmania parasites in diverse geographic regions using RAPD in order to constitute a pool of PCR targets having the potential to differentiate among the VL parasites. 100 different oligonucleotide decamers having arbitrary DNA sequences were screened for reproducible amplification and a selection of 28 was used to amplify DNA from 12 L. donovani, L. archibaldi and L. infantum strains having diverse origins. A total of 155 bands were amplified of which 60.65% appeared polymorphic. 7 out of 28 primers provided monomorphic patterns. Phenetic analysis allowed clustering the parasites according to their geographical origin. Differentially amplified bands were selected, among them 22 RAPD products were successfully cloned and sequenced. Bioinformatic analysis allowed mapping of the markers and sequences and priming sites analysis. This study was complemented with Southern-blot to confirm assignment of markers to the kDNA. The bioinformatic analysis identified 16 nuclear and 3 minicircle markers. Analysis of these markers highlighted polymorphisms at RAPD priming sites with mainly 59 end transversions, and presence of inter- and intra-taxonomic complex sequence and microsatellites variations; a bias in transitions over transversions and indels between the different sequences compared is observed, which is however less marked between L. infantum and L. donovani. The study delivers a pool of well-documented polymorphic DNA markers, to develop molecular diagnostics assays to characterize and differentiate VL causing agents.
PY 2014
PD OCT
SO Plos One
SN 1932-6203
PU Public Library Science
VL 9
IS 10
UT 000343662500076
DI 10.1371/journal.pone.0109773
ID 38559
ER

EF