通过动质体DNA的杂交和限制性内切酶消化来检测和鉴定来自哺乳动物及传播媒介的利什曼原虫种类和菌株。

Detection and characterization of Leishmania species and strains from mammals and vectors by hybridization and restriction endonuclease digestion of kinetoplast DNA.

作者信息

Jackson P R, Lawrie J M, Stiteler J M, Hawkins D W, Wohlhieter J A, Rowton E D

出版信息

Vet Parasitol. 1986 Mar;20(1-3):195-215. doi: 10.1016/0304-4017(86)90100-7.

Abstract

Leishmania parasites from animals, man or insect vectors were characterized by the gel electrophoresis of restriction endonuclease enzyme-produced mitochondrial (kinetoplast) DNA (kDNA) fragments and/or by DNA-DNA hybridization with 32P-labelled cloned, or uncloned, kDNA fragment probes from type isolates. The electrophoretic separation of kDNA fragments is a sensitive method for detecting genetic similarities and differences among Leishmania. Parasites with similar kDNA restriction fragment patterns belong to the same schizodeme and schizodeme analysis is useful for studying Leishmania populations. Cloned, species-specific kDNA probes detected Leishmania in sandflies and in liver, spleen or blood preparations from infected animals. Cloned DNA probes also hybridized to immobilized kDNA from in vitro cultivated promastigotes and detected as few as 100 parasites in a species-specific manner. Sensitive DNA hybridization probes should be useful in research on the immunology, chemotherapy or epidemiology of animal and human leishmaniasis.

摘要

来自动物、人类或昆虫媒介的利什曼原虫寄生虫，通过限制性内切酶产生的线粒体（动基体）DNA（kDNA）片段的凝胶电泳和/或与来自标准株的32P标记的克隆或未克隆的kDNA片段探针进行DNA-DNA杂交来进行特征鉴定。kDNA片段的电泳分离是检测利什曼原虫之间遗传相似性和差异的灵敏方法。具有相似kDNA限制性片段模式的寄生虫属于同一裂殖体型，裂殖体型分析对于研究利什曼原虫种群很有用。克隆的、种特异性的kDNA探针可检测白蛉以及感染动物的肝脏、脾脏或血液制剂中的利什曼原虫。克隆的DNA探针也能与体外培养的前鞭毛体固定化的kDNA杂交，并以种特异性方式检测到低至100个寄生虫。灵敏的DNA杂交探针在动物和人类利什曼病的免疫学、化疗或流行病学研究中应会很有用。