DETECTION AND CLASSIFICATION OF TRYPANOSOMA-CRUZI BY DNA HYBRIDIZATION WITH NONRADIOACTIVE PROBES

被引:16
|
作者
SOLARI, A [1 ]
VENEGAS, J [1 ]
GONZALEZ, E [1 ]
VASQUEZ, C [1 ]
机构
[1] UNIV TALCA,FAC RECURSOS NAT,DEPT CIENCIAS BIOL,TALCA 747,CHILE
来源
JOURNAL OF PROTOZOOLOGY | 1991年 / 38卷 / 06期
关键词
BIOTINYLATED PROBES; KINETOPLAST DNA;
D O I
10.1111/j.1550-7408.1991.tb06080.x
中图分类号
Q95 [动物学];
学科分类号
071002 ;
摘要
Total or kinetoplast DNA (kDNA) from 72 isolates and clones of Trypanosoma cruzi as well as from nine related trypanosomatids were analyzed by dot hybridization using nonradioactive kDNA or cloned minicircle fragments as probes. Biotinylated-kDNA probes generated by nick-translation proved reliable for distinguishing Zymodeme 1 and Zymodeme 2bol of T. cruzi parasites. In contrast, digoxigenin-labeled kDNA obtained by random-priming did not distinguish among T. cruzi isolates but did distinguish among New World leishmanias. Cloned minicircle fragments labeled with digoxigenin gave the same results as digoxigenin-labeled kDNA, except for a 10-fold decrease in sensitivity. Digoxigenin-labeled DNA probes proved useful in unambiguously detecting T. cruzi from different geographic regions of America. However, T. rangeli and T. cruzi marinkellei were not distinguished by these probes.
引用
收藏
页码:559 / 565
页数:7
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