Comparison of molecular markers for strain typing of Leishmania infantum

被引:70
作者
Botilde, Yanick
Laurent, Thierry
Tintaya, Wilber Quispe
Chicharro, Carmen
Canavate, Carmen
Cruz, Israel
Kuhls, Katrin
Schoenian, Gabriele
Dujardin, Jean-Claude
机构
[1] Inst Trop Geneeskunde, Mol Parasitol Lab, B-2000 Antwerp, Belgium
[2] Inst Salud Carlos III, Natl Ctr Microbiol, Dept Parasitol, WHO Collaborating Ctr Leishmaniasis, Majadahonda 28220, Spain
[3] Charite Univ Med Berlin, Inst Mikrobiol & Hyg, D-10098 Berlin, Germany
关键词
Leishmania infantum; MON-1; Spain; PCR-RFLP; RAPD; microsatellites;
D O I
10.1016/j.meegid.2006.02.003
中图分类号
R51 [传染病];
学科分类号
100401 ;
摘要
The epidemiology of Leishmania infantum, the etiological agent of visceral leishmaniasis, is changing rapidly; hence powerful typing tools are required in order to monitor the parasite populations spreading and to adapt adequate control measures. We compared here the resolving power of four molecular methods at the zymodeme level: PCR-RFLP analysis of kDNA minicircles (kDNAPCR-RFLP) and antigen genes (cysteine proteinase b and major surface protease, cpb- and gp63PCR-RFLP), multilocus microsatellite typing (MLMT) and random amplification of polymorphic DNA (RAPD) were applied to samples of 25 L. infantum MON-1 strains obtained from different hosts (HIV+ patients, HIV-patients and dogs) coming from three Spanish foci: Madrid, Mallorca and Ibiza. While RAPD was not sufficiently resolving, the other three methods allowed genotyping within the zymodeme. KDNAPCR-RFLP and MLMT were the most discriminatory and appeared the most adequate for strain fingerprinting. In an eco-geographical context, cpbPCR-RFLP, MLMT and kDNAPCR-RFLP were all informative: they showed here a similar picture, with the existence of cluster(s) of isolates from the islands and other one(s) of mixed composition (Madrid and the islands). None of the markers revealed an association with the host type or the clinical form. In general, there was a significant correlation between each pair of distances calculated from the cpb, microsatellite and kDNA data, respectively, but visual inspection of the trees revealed a better congruence between cpb and microsatellite trees. The methods used here are complementary and each adapted to answer specific epidemiological questions. Their choice should be the result of a compromise between the required resolving power, the genetic features of the respective markers and the technical aspects. (c) 2006 Elsevier B.V. All rights reserved.
引用
收藏
页码:440 / 446
页数:7
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