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首页> 外文期刊>Journal of Parasitology >Discrimination of Leishmania braziliensis variants by kDNA signatures produced by LSSP-PCR.
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Discrimination of Leishmania braziliensis variants by kDNA signatures produced by LSSP-PCR.

机译：通过LSSP-PCR产生的kDNA签名区分巴西利什曼原虫变种。

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The conventional methods for identification and typing of Leishmania species depend on previous culture isolation of the parasites. Not infrequently, culture is unsuccessful and may result in misrepresentation of the heterogeneity of the original isolate. Thus, more reliable and precise identification of genotypes of Leishmania spp. is important for a better clinical and epidemiological understanding of the disease. We evaluated the potential of LSSP-PCR targeting kDNA minicircles in discriminating different variants of the parasite with the use of clinical samples directly or cultivated parasites. The 1st step of this procedure consists of the amplification of the minicircles by conventional PCR; the 2nd step is low-stringency amplification of the minicircles previously amplified, with the use of 1 of the primers. Although LSSP-PCR produced complex and distinct kDNA signatures for isolates representing different species, further experiments demonstrated that the approach had the potential for discriminating intraspecific variants of L. braziliensis. Thus, the generated profiles were too variable to be useful as markers for species identification. Moreover, we demonstrated that the approach can be directly applied to clinical samples. In conclusion, LSSP-PCR targeting kDNA minicircles produces profiles that reflect polymorphisms of the predominant classes of minicircles, and can be useful for studies aimed at discriminating Leishmania braziliensis genotypes without the need for previous cultivation of the parasite.
机译：用于鉴定和分类利什曼原虫种类的常规方法取决于先前寄生虫的培养分离。并非总是如此，培养是不成功的，并且可能导致原始分离物异质性的错误表述。因此，更可靠和精确地鉴定利什曼原虫属的基因型。对于更好地对该疾病的临床和流行病学理解至关重要。我们通过直接使用临床样品或培养的寄生虫，评估了针对kDNA微小环的LSSP-PCR的潜力，以区分该寄生虫的不同变体。该步骤的第一步是通过常规PCR扩增小圆;第二步是使用一种引物对先前扩增的小圆进行低严格度扩增。尽管LSSP-PCR为代表不同物种的分离株产生了复杂而独特的kDNA标记，但进一步的实验表明该方法具有区分巴西乳杆菌种内变异的潜力。因此，生成的轮廓太可变而不能用作物种识别的标记。此外，我们证明了该方法可以直接应用于临床样品。总之，针对kDNA小圆环的LSSP-PCR产生的图谱反映了主要小圆环类的多态性，可用于旨在区分巴西利什曼原虫基因型的研究，而无需先前的寄生虫培养。

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