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基于动基体DNA微小环的伊氏锥虫(锥虫亚属)特异性探针。

Specific probes for Trypanosoma (Trypanozoon) evansi based on kinetoplast DNA minicircles.

作者信息

Masiga D K, Gibson W C

机构信息

Department of Pathology, School of Veterinary Science, University of Bristol, Langford, U.K.

出版信息

Mol Biochem Parasitol. 1990 May;40(2):279-83. doi: 10.1016/0166-6851(90)90049-r.

Abstract

Trypanosoma evansi is difficult to distinguish from other members of subgenus Trypanozoon, save for its inability to develop cyclically in the tsetse fly and its characteristic kinetoplast DNA (kDNA). We have used cloned kDNA minicircle fragments as specific probes to distinguish T. evansi from other trypanosomes of subgenus Trypanozoon. Two probes were required, each specific for one of the subgroups of T. evansi previously described. Probe A reacted only with the major isoenzyme group of T. evansi stocks, which have minicircle type A and occur in South America, Kenya, Sudan, Nigeria and Kuwait. The probe did not hybridise with various Trypanosoma brucei spp. stocks, Trypanosoma vivax, Trypanosoma congolense or Trypanosoma simiae, nor with trypanosomes of the minor isoenzyme group of T. evansi stocks found in Kenya with type B minicircles. Probe B was specific for the latter. The probes were sensitive down to a level of 100 trypanosomes in a dot blot. These probes thus provide a simple means of distinguishing T. evansi from T. brucei spp. using comparatively few trypanosomes and without resort to tsetse transmission experiments.

摘要

伊氏锥虫很难与布氏锥虫亚属的其他成员区分开来,只是它无法在采采蝇体内进行循环发育,且具有独特的动基体DNA(kDNA)。我们使用克隆的kDNA微小环片段作为特异性探针,以区分伊氏锥虫与布氏锥虫亚属的其他锥虫。需要两种探针,每种探针针对先前描述的伊氏锥虫的一个亚群具有特异性。探针A仅与伊氏锥虫虫株的主要同工酶组发生反应,这些虫株具有微小环A型,分布于南美洲、肯尼亚、苏丹、尼日利亚和科威特。该探针不与各种布氏锥虫虫株、活泼锥虫、刚果锥虫或西氏锥虫杂交,也不与在肯尼亚发现的具有B型微小环的伊氏锥虫虫株的次要同工酶组的锥虫杂交。探针B对后者具有特异性。在斑点印迹中,这些探针的灵敏度低至100个锥虫的水平。因此,这些探针提供了一种简单的方法,可使用相对较少的锥虫来区分伊氏锥虫和布氏锥虫,而无需进行采采蝇传播实验。

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