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首页> 外文期刊>The American Journal of Tropical Medicine and Hygiene >Molecular Identification of Leishmania spp. DNA from Archived Giemsa-Stained Slides of Patients from Salta, Argentina
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Molecular Identification of Leishmania spp. DNA from Archived Giemsa-Stained Slides of Patients from Salta, Argentina

机译：Leishmania SPP的分子鉴定。 来自阿根廷萨尔塔患者的存档Giemsa染色幻灯片的DNA

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Cutaneous leishmaniasis is endemic in Salta province, which belongs to the northwest of Argentina. Leishmania spp. DNA from Giemsa-stained slides of up to 12 years in storage of patients from Salta was characterized through polymerase chain reaction (PCR) restriction fragment length polymorphism (RFLP). One hundred smears positive for microscopy, classified in a semiquantitative scale for amastigote density, were analyzed. Also, Leishmanin skin test (LST) results were included. DNA extraction was carried out applying lysis buffer with proteinase K, and then DNA was amplified with ribosomal internal transcribed spacer 1 primers. PCR products were digested with HaeIII enzyme. All PCR-positive smears (74/100) belonged to Viannia subgenus. A statistically significant, directly proportional relationship between semiquantitative microscopy and PCR results was detected. All patients had LST-positive results (induration = 5 mm), and the smears of those with smaller induration (LST 19 mm) gave a higher proportion of positive PCR results. This study determined that smear age did not affect PCR positivity, which allows retrospective analyzes and suggests smears might be useful for molecular complementary diagnosis. Because Leishmania (Viannia) braziliensis is the main circulating species in the study area, determining Viannia subgenus in all analyzed samples confirms previous findings. PCR positivity showed statistically significant differences according to semiquantitative microscopy, highlighting the importance of parasite burden in the diagnostic sensitivity of the method. Considering that smears of patients with smaller LST induration were more positive in PCR, a negative smear from patients with positive LST response, but 19 mm, could actually represent a false-negative result.
机译：皮肤利什曼病是萨尔塔省的地方，属于阿根廷西北部。 Leishmania SPP。通过聚合酶链反应（PCR）限制性片段长度多态性（RFLP），表征来自盐酸患者的Giemsa染色载体的DNA高达12年的储存。分析了一百个涂片的显微镜阳性，分析了以氨神经奇密度的半定量规模分类。此外，包括Leishmanin皮肤测试（LST）结果。通过蛋白酶K进行DNA萃取施用裂解缓冲液，然后用核糖体内转录的间隔物1引物扩增DNA。用HAEIII酶消化PCR产物。所有PCR阳性涂片（74/100）属于Viannnia subguus。检测到半定位显微镜和PCR结果之间的统计学意义，直接比例的关系。所有患者均具有LST-阳性结果（硬结＆ = 5mm），并且较小（LST <19mm）较小的那些涂片的涂片产生较高的阳性PCR结果。该研究确定了涂抹年龄不影响PCR阳性，这允许回顾性分析，并表明涂片可能对分子互补诊断有用。由于利什曼尼亚（viannia）巴西是研究区的主要循环物种，因此在所有分析的样品中确定viannia子属亚血症证实了以前的研究结果。 PCR阳性根据半定位显微镜显示出统计学意义差异，突出了寄生虫负荷在该方法的诊断敏感性中的重要性。考虑到LST硬化较小的患者的涂片在PCR中更阳性，来自阳性LST反应的患者的阴性涂片，但是＆ 19毫米，实际上可以代表一个假阴性结果。

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