Malaria y Protozoos Emergentes, Servicio de Parasitología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Carretera Majadahonda-Pozuelo km 2, Majadahonda, Madrid, Spain.
Mem Inst Oswaldo Cruz. 2010 Sep;105(6):823-8. doi: 10.1590/s0074-02762010000600016.
We present filaria-nested polymerase chain reaction (PCR), which is based on amplification of first internal transcribed spacer rDNA to distinguish three parasitic filarial species (Onchocerca volvulus, Mansonella ozzardi and Mansonella perstans) that can be found in the Amazon Region. Nested PCR-based identifications yielded the same results as those utilizing morphological characters. Nested PCR is highly sensitive and specific and it detects low-level infections in both humans and vectors. No cross-amplifications were observed with various other blood parasites and no false-positive results were obtained with the nested PCR. The method works efficiently with whole-blood, blood-spot and skin biopsy samples. Our method may thus be suitable for assessing the efficacy of filaria control programmes in Amazonia by recording parasite infections in both the human host and the vector. By specifically differentiating the major sympatric species of filaria, this technique could also enhance epidemiological research in the region.
我们提出了内套式聚合酶链反应（PCR），它基于第一内部转录间隔区 rDNA 的扩增，以区分在亚马逊地区发现的三种寄生丝虫（盘尾丝虫、奥氏曼森线虫和曼氏裂头蚴）。基于内套式 PCR 的鉴定结果与利用形态特征的结果相同。内套式 PCR 具有高度的敏感性和特异性，可检测人类和媒介中低水平的感染。与各种其他血液寄生虫没有交叉扩增，并且内套式 PCR 没有假阳性结果。该方法在全血、血斑和皮肤活检样本上都能有效地工作。因此，我们的方法可以通过记录人类宿主和媒介中的寄生虫感染来评估亚马逊地区的丝虫控制计划的效果。通过特异性地区分主要共生的丝虫种类，该技术还可以增强该地区的流行病学研究。
