通过“斑点”杂交快速鉴定克氏锥虫分离株

Rapid identification of Trypanosoma cruzi isolates by 'dot-spot' hybridization.

作者信息

Sanchez D O, Madrid R, Engel J C, Frasch A C

出版信息

FEBS Lett. 1984 Mar 12;168(1):139-42. doi: 10.1016/0014-5793(84)80223-9.

Abstract

The presence of isolate-specific Trypanosoma cruzi minicircles has been shown in the kinetoplast DNA of this parasite. This led to the rapid identification of isolates and clones of trypanosomes by means of 'dot-spot' hybridizations with molecularly cloned minicircle probes. Unexpectedly, whole kDNAs were also suitable as probes for this purpose, provided that filters were washed under stringent conditions. This was attributed to the presence of the above-mentioned isolate-specific minicircle sequences. The fact that parasites could be directly spotted onto nitrocellulose filters simplified the rapid routine screening of a large number of samples.

摘要

已在这种寄生虫的动基体DNA中发现了特定分离株的克氏锥虫微小环。这使得通过与分子克隆的微小环探针进行“点斑”杂交，能够快速鉴定锥虫的分离株和克隆。出乎意料的是，只要在严格条件下洗涤滤膜，整个动基体DNA也适合用作此目的的探针。这归因于上述特定分离株微小环序列的存在。寄生虫可直接点样到硝酸纤维素滤膜上这一事实简化了对大量样品的快速常规筛选。