Comparison of molecular markers for strain typing of Leishmania infantum
Comparison of molecular markers for strain typing of Leishmania infantum
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DOI：
10.1016/j.meegid.2006.02.003
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发表时间：
2006-11-01
影响因子：
3.2
通讯作者：
Dujardin, Jean-Claude
中科院分区：
文献类型：
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作者：
Botilde, Yanick;Laurent, Thierry;Dujardin, Jean-Claude
婴儿利什曼原虫是内脏利什曼病的病原体，其流行病学正在迅速变化；因此，需要强大的分型工具来监测寄生虫种群的传播并采取适当的控制措施。我们比较了4种分子方法在酶谱水平上的分辨能力：kDNA小环的PCR-RFLP分析(kDNAPCR-RFLP)和抗原基因(半胱氨酸蛋白酶b和主要表面蛋白酶，cpb-和gp63PCR-RFLP)、多位点微卫星分型(MLMT)和随机扩增多态性DNA(RAPD)方法对来自西班牙马德里、马略卡岛和伊比沙岛的25株不同宿主(HIV+患者、HIV患者和狗)的L.infantum mon1菌株进行了检测。在不能完全分辨的情况下，其他三种方法都可以在酵母菌体内进行基因分型。KDNAPCR-RFLP和MLMT最具区分性，似乎最适合菌株指纹图谱。在生态地理背景下，cpbPCR-RFLP、MLMT和kDNAPCR-RFLP都是信息性的：它们在这里显示了相似的图像，存在来自岛屿的菌株群(S)和另一群混合组成的分离物(S)(马德里和岛屿)。没有一个标记物显示与宿主类型或临床形式有关。总的来说，分别从CPB、微卫星和kDNA数据计算出的每对距离之间存在显著的相关性，但对这些树的目测检查显示，CPB和微卫星树之间的一致性更好。这里使用的方法是相辅相成的，每种方法都适用于回答特定的流行病学问题。他们的选择应该是在所需的分辨率、各自标记的遗传特征和技术方面之间妥协的结果。(C)2006爱思唯尔B.V.保留所有权利。
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.