沙特阿拉伯西部麦地那省沙蝇中的利什曼原虫感染的分子特征。
Molecular characterization of Leishmania infection in sand flies from Al-madinah Al-munawarah province, western Saudi Arabia.
机构信息
Medical Laboratories Technology Department, Faculty of Applied Medical Sciences, Taibah University, Al-madinah Al-munawarah, PO Box 30001, Saudi Arabia.
出版信息
Exp Parasitol. 2013 Jun;134(2):211-5. doi: 10.1016/j.exppara.2013.02.016. Epub 2013 Mar 6.
Cutaneous leishmaniasis (CL) is caused by various species of the genus Leishmania. The disease is considered a major health problem in different areas of Saudi Arabia including Al-madinah Al-munawarah province. We aimed to identify Leishmania species isolated from sand fly vectors by molecular analysis. Sand fly sampling was carried out from May 2010 to October 2010 in province of Al-madinah Al-munawarah from four different localities. Female sand flies collected were subjected to DNA extraction followed by molecular analysis using the semi-nested PCR and conventional PCR protocols, respectively, against minicircle kDNA and ribosomal internal transcribed spacer 1 (ITS1-rDNA). The PCR positive specimens against ITS1-rDNA locus were digested for further confirmation of species identification. A total of 2910 sand flies were collected. Phlebotomus papatasi accounted for 93.8% (1673 males and 1057 females), however, the number of Phlebotomus sergenti was only 180 (109 males and 71 females). Sixty-two out of 250 (23.7%) female P. papatasi tested for Leishmania parasite were positive for Leishmania major using the semi-nested PCR method against kDNA. All of the 62 positive specimens produced a band size 650 bp. A 31% of female P. sergenti were positive against kDNA of Leishmania tropica and produced a 720 bp band. These positive P. sergenti for L. tropica DNA produced ITS1-PCR-RFLP profile showed two bands of ∼200 bp and 57 bp which are specific for L. tropica, confirming the presence of L. tropica in P. sergenti. However, the ITS1-PCR-RFLP profile showed two bands of ∼203 bp and 132 bp which are specific for L. major in P. papatasi. We concluded that, the semi-nested PCR method against kDNA and the ITS1-PCR-RFLP analysis are useful tools for molecular identification of both L. major and L. tropica. A multicenter study is necessary in order to evaluate the extent of the disease and functional analysis of new Leishmania genes.
皮肤利什曼病（CL）是由利什曼属的各种物种引起的。该疾病被认为是沙特阿拉伯不同地区的一个主要健康问题，包括麦地那省。我们的目的是通过分子分析确定从沙蝇媒介中分离出的利什曼菌种。沙蝇采样于 2010 年 5 月至 10 月在麦地那省的四个不同地点进行。采集的雌性沙蝇进行 DNA 提取，然后分别使用半巢式 PCR 和常规 PCR 方案对小型环 kDNA 和核糖体内部转录间隔区 1（ITS1-rDNA）进行分子分析。针对 ITS1-rDNA 基因座的 PCR 阳性标本进行消化，以进一步确认种属鉴定。共采集 2910 只沙蝇。斑虻属（Phlebotomus）占 93.8%（1673 只雄性和 1057 只雌性），然而，仅采集到 180 只白蛉属（Phlebotomus）（109 只雄性和 71 只雌性）。用针对 kDNA 的半巢式 PCR 方法检测 250 只雌性斑虻属（Phlebotomus）中的 62 只利什曼原虫寄生虫呈阳性，其中 62 只均为利什曼原虫（Leishmania）大种。所有 62 个阳性标本均产生 650 bp 大小的条带。31%的雌性白蛉属（Phlebotomus）对利什曼原虫热带种的 kDNA 呈阳性，并产生 720 bp 的条带。这些对白蛉属（Phlebotomus）热带种 DNA 呈阳性的白蛉属（Phlebotomus）产生的 ITS1-PCR-RFLP 图谱显示出两条约 200 bp 和 57 bp 的带，这是利什曼原虫热带种的特异性带，证实了白蛉属（Phlebotomus）中存在利什曼原虫热带种。然而，在斑虻属（Phlebotomus）中，ITS1-PCR-RFLP 图谱显示出两条约 203 bp 和 132 bp 的带，这是利什曼原虫大种的特异性带。我们得出结论，针对 kDNA 的半巢式 PCR 方法和 ITS1-PCR-RFLP 分析是鉴定利什曼原虫大种和利什曼原虫热带种的有用工具。有必要进行多中心研究，以评估疾病的程度和新利什曼原虫基因的功能分析。
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