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首页> 外文期刊>Parasitology >Sensitive diagnosis of cutaneous leishmaniasis by lesion swab sampling coupled to qPCR.
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Sensitive diagnosis of cutaneous leishmaniasis by lesion swab sampling coupled to qPCR.

机译：病变拭子抽样敏感诊断皮肤乳山生成耦合到QPCR。

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摘要

Variation in clinical accuracy of molecular diagnostic methods for cutaneous leishmaniasis (CL) is commonly observed depending on the sample source, the method of DNA recovery and the molecular test. Few attempts have been made to compare these variables. Two swab and aspirate samples from lesions of patients with suspected CL (n = 105) were evaluated alongside standard diagnosis by microscopic detection of amastigotes or culture of parasites from lesion material. Three DNA extraction methods were compared: Qiagen on swab and aspirate specimens, Isohelix on swabs and Boil/Spin of lesion aspirates. Recovery of Leishmania DNA was evaluated for each sample type by real-time polymerase chain reaction detection of parasitic 18S rDNA, and the diagnostic accuracy of the molecular method determined. Swab sampling combined with Qiagen DNA extraction was the most efficient recovery method for Leishmania DNA, and was the most sensitive (98%; 95% CI: 91-100%) and specific (84%; 95% CI: 64-95%) approach. Aspirated material was less sensitive at 80% (95% CI: 70-88%) and 61% (95% CI: 50-72%) when coupled to Qiagen or Boil-Spin DNA extraction, respectively. Swab sampling of lesions was painless, simple to perform and coupled with standardized DNA extraction enhances the feasibility of molecular diagnosis of CL.
机译：临床准确性的临床准确性，其分子诊断方法对皮肤LeishManiaisis（CL）的临床准确性，根据样品源，DNA回收率和分子试验的方法观察。已经尝试了很少的尝试来比较这些变量。通过微观检测来自病变材料的寄生虫的微观检测，评估来自疑似Cl（n = 105）的患者病变的两种拭子和吸气样品。比较了三种DNA提取方法：在拭子和吸气标本上进行QIAGEN，ISOHELIX在拭子上并煮沸/旋转病变吸气。通过寄生18SRDNA的实时聚合酶链反应检测对每个样品类型评估LeishMania DNA的回收，并确定分子方法的诊断精度。拭子抽样结合QIAGEN DNA提取是LeishMania DNA最有效的回收方法，是最敏感的（98％; 95％CI：91-100％）和特异性（84％; 95％CI：64-95％）方法。当偶联到QIAGEN或沸腾DNA提取时，吸气材料在80％（95％CI：70-88％）和61％（95％CI：50-72％）下敏感。拭子的病变抽样是无痛的，简单的表演和结合标准化DNA提取，提高了CL的分子诊断的可行性。

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