Health Sciences Postgraduate Program-PCS, Universidade Estadual de Maringá, Maringá, State of Paraná, Brazil.
PLoS One. 2013 Jul 5;8(7):e62473. doi: 10.1371/journal.pone.0062473. eCollection 2013.
Cutaneous leishmaniasis (CL) can occur in skin and mucosa, causing disfiguring lesions. The laboratory diagnosis of CL involves immunological methods and optical detection of the parasite, al of which have limitations. There is a need for more effective diagnostic methods for CL which wil allow treatment to be initiated more promptly in order to help prevent the development of severe forms of mucosal disease, and to estimate the prognosis of the infection. The polymerase chain reaction (PCR) has been widely used to diagnose CL, because of its higher sensitivity. This study estimated the accuracy and compared PCRs of samples from lesion scarification (PCR-L) and blood sample-enriched leukocytes (PCR-B) with three conventional diagnostic techniques: parasite direct search (DS), Montenegro skin test (MST), and indirect immunofluorescence reaction (IIF). The study included 276 patients under suspicion of CL. We conducted a cross-sectional study, in which patients were selected by convenience sampling. We used MP3H/MP1L primers to generate a Leishmania (Viannia) (minicircle kDNA) fragment of 70-bp. Of 106 patients with CL, 83.87%, 51.67%, 64.52%, 85.71%, or 96.10% tested positive by PCR-L, PCR-B, DS, IIF, or MST, respectively. Five patients tested positive only by PCR-L, and two other patients only by PCR-B. PCR-L is indicated for use in patients with chronic lesions or Leishmania reinfection, which may progress to mucosal lesion. PCR-B is indicated for use in patients with negative results in conventional tests or for patients with no apparent lesion. PCR is not only useful in diagnosing CL but also helps to identify the infecting species.
皮肤利什曼病(CL)可发生于皮肤和黏膜,导致毁容性病变。CL 的实验室诊断包括免疫学方法和寄生虫光学检测,但都存在局限性。因此,CL 需要更有效的诊断方法,以便更及时地开始治疗,从而有助于防止黏膜疾病的严重形式发展,并估计感染的预后。聚合酶链反应(PCR)已广泛用于 CL 的诊断,因为它具有更高的灵敏度。本研究评估了 PCR 的准确性,并比较了病变划痕(PCR-L)和血液白细胞富集样本(PCR-B)的 PCR 与三种常规诊断技术:寄生虫直接搜索(DS)、曼托瓦尼皮肤试验(MST)和间接免疫荧光反应(IIF)。该研究纳入了 276 例疑似 CL 的患者。我们进行了一项横断面研究,通过便利抽样选择患者。我们使用 MP3H/MP1L 引物生成 70-bp 的利什曼原虫(Viannia)(微环 kDNA)片段。在 106 例 CL 患者中,PCR-L、PCR-B、DS、IIF 或 MST 的阳性率分别为 83.87%、51.67%、64.52%、85.71%或 96.10%。5 例患者仅 PCR-L 阳性,另有 2 例患者仅 PCR-B 阳性。PCR-L 适用于慢性病变或利什曼原虫再感染的患者,后者可能进展为黏膜病变。PCR-B 适用于常规检测结果阴性或无明显病变的患者。PCR 不仅有助于诊断 CL,还有助于鉴定感染物种。