Toé L, Back C, Adjami A G, Tang J M, Unnasch T R
Onchocerciasis Control Programme, Bouake, Côte d'Ivoire.
Bull World Health Organ. 1997;75(5):443-7.
In recent years, methods for the identification of the filarial worm Onchocerca volvulus and its vector, blackflies of the Simulium damnosum complex (S. damnosum sensu lato (s.l.)), based on the amplification of parasite and vector DNA sequences with the polymerase chain reaction (PCR), have been developed. Routine application of these methods requires techniques for sample collection and preservation that are compatible with the limitations of field collection, yet preserve DNA in a form suitable for PCR. Two different methods for sample preservation were evaluated by the field collection teams and the DNA probe laboratory of the Onchocerciasis Control Programme in West Africa. The most successful involved the preservation of material from O. volvulus and its associated vectors in a dried state on microscope slides. Of over 1200 parasite samples preserved in this manner, more than 93% retained DNA yielding positive results in PCR analysis (1208/1291). Vector material (malpighian tubules and ovaries) preserved in the same manner on the same microscope slides also yielded DNA that was suitable for PCR.
近年来,基于用聚合酶链反应(PCR)扩增寄生虫和媒介DNA序列,已开发出鉴定盘尾丝虫及其媒介——恶蚋复合体(狭义的恶蚋)黑蝇的方法。这些方法的常规应用需要与野外采集的局限性相适应的样本采集和保存技术,同时还要以适合PCR的形式保存DNA。西非盘尾丝虫病控制项目的野外采集团队和DNA探针实验室评估了两种不同的样本保存方法。最成功的方法是将来自盘尾丝虫及其相关媒介的材料以干燥状态保存在载玻片上。以这种方式保存的1200多个寄生虫样本中,超过93%的样本保留了能在PCR分析中产生阳性结果的DNA(1208/1291)。以同样方式保存在同一张载玻片上的媒介材料(马氏管和卵巢)也产生了适合PCR的DNA。