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利用碱基特异性荧光染料和判别分析对克氏锥虫株进行特征描述和分组

Characterization and grouping of Trypanosoma cruzi stocks by DNA base-specific fluorochromes and discriminant analysis.

作者信息

Mühlpfordt H, Berger J

机构信息

Abteilung für Protozoologie, Bernhard-Nocht-Institut für Tropenmedizin, Hamburg, Federal Republic of Germany.

出版信息

Parasitol Res. 1990;76(4):319-25. doi: 10.1007/BF00928186.

Abstract

Fluorochromes with G-C and A-T specificity were used for a single-cell DNA analysis of the blood-stream forms of 14 Trypanosoma cruzi stocks in a cytofluorometric assay. The kinetoplast contained 22.3%-37.9% of the total DNA G-C base content and 42.7%-63.5% of the total DNA A-T base content. In spite of these differences, the mean base A-T/G-C ratio of the total DNA was 1.11 and was nearly constant in all stocks. The G-C base ratio of kinetoplast/nucleus resulted in a grouping corresponding with the peanut agglutinin (PNA)- and wheat germ agglutinin (WGA)-type characteristics of the T. cruzi stocks. The discriminant analysis revealed relationships, in that each stock contained some trypanosomes with DNA fluorescence characteristics of common to at least one other stock. After chromomycin A3 staining, the mean hit rates for the classification into group 1 PNA and the WGA group were 99% and 96%, respectively, and the respective rates obtained after DAPI application were 84% and 94%.

摘要

在细胞荧光分析中,使用具有G-C和A-T特异性的荧光染料对14株克氏锥虫血流型进行单细胞DNA分析。动基体含有总DNA中G-C碱基含量的22.3%-37.9%和总DNA中A-T碱基含量的42.7%-63.5%。尽管存在这些差异,但总DNA的平均A-T/G-C碱基比为1.11,且在所有株系中几乎恒定。动基体/细胞核的G-C碱基比导致了一种分组,这与克氏锥虫株系的花生凝集素(PNA)和麦胚凝集素(WGA)类型特征相对应。判别分析揭示了其中的关系,即每个株系都包含一些具有至少与其他一个株系共有的DNA荧光特征的锥虫。经放线菌素A3染色后,分类为1组PNA和WGA组的平均命中率分别为99%和96%,应用4',6-二脒基-2-苯基吲哚(DAPI)后获得的相应比率分别为84%和94%。

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