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首页> 外文期刊>Journal of Clinical Microbiology >Diagnosis of cutaneous leishmaniasis and species discrimination of parasites by PCR and hybridization.
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Diagnosis of cutaneous leishmaniasis and species discrimination of parasites by PCR and hybridization.

机译：PCR和杂交技术诊断皮肤利什曼病和寄生虫的种类。

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The aim of this study was to assess the efficacy of PCR methodology in establishing the diagnosis of cutaneous leishmaniasis in patients from areas of endemicity in Venezuela. Biopsies from 233 patients with cutaneous ulcers suggestive of leishmaniasis were analyzed by PCR, employing oligonucleotides directed against conserved regions of kinetoplast DNA (kDNA), and the PCR products were then hybridized to nonradioactively labeled, species-specific, cloned kDNA fragments. The ability of PCR to detect Leishmania cells was compared with those of the conventional methodologies: skin testing with killed promastigotes (Montenegro test), examination of Giemsa-stained biopsy smears, and in vitro culture of biopsy tissue. The PCR-hybridization technique detected the presence of Leishmania cells in 98% of patients clinically diagnosed as having leishmaniasis and also positive by the Montenegro skin test. In comparison, leishmania positivity was found in only 42% of cultures and 64% of biopsy smears. By hybridizing the PCR product to new kDNA probes specific for either Leishmania mexicana or Leishmania braziliensis, we found that both species are major causes of cutaneous leishmaniasis in Venezuela, and the species identification was confirmed by restriction enzyme analysis of kDNA from biopsy cultures. This work demonstrates that PCR coupled with hybridization is useful not only for the diagnosis of cutaneous leishmaniasis but also for the taxonomic discrimination essential for both epidemiology and therapy. This technique can be used to diagnose leishmaniasis in a country in which the disease is endemic and can perhaps be adapted for use in a rural clinic.
机译：这项研究的目的是评估PCR方法在建立委内瑞拉流行地区患者皮肤利什曼病诊断中的功效。通过PCR分析233例提示利什曼病的皮肤溃疡患者的活检，采用针对运动塑料DNA（kDNA）保守区的寡核苷酸，然后将PCR产物与非放射性标记的，物种特异性的克隆的kDNA片段杂交。将PCR检测利什曼原虫细胞的能力与常规方法进行了比较：使用杀死的前鞭毛体进行皮肤测试（黑山测试），检查吉姆萨染色的活检涂片以及活检组织的体外培养。 PCR杂交技术在98％的临床诊断为患有利什曼病且黑山皮肤试验也呈阳性的患者中检测到利什曼原虫细胞的存在。相比之下，仅42％的培养物和64％的活检涂片发现利什曼原虫阳性。通过将PCR产物与特异于墨西哥利什曼原虫或巴西利什曼原虫的新kDNA探针杂交，我们发现这两个物种都是委内瑞拉皮肤利什曼原虫病的主要原因，并且通过对活检培养物中的kDNA进行限制性酶切分析证实了物种鉴定。这项工作表明，PCR与杂交结合不仅可用于诊断皮肤利什曼病，而且可用于流行病学和治疗必不可少的分类学鉴别。该技术可用于在该病流行的国家诊断利什曼病，并且可能适合在乡村诊所中使用。

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