Murthy V K, Dibbern K M, Campbell D A
Department of Microbiology and Immunology, University of California, Los Angeles 90024.
Mol Cell Probes. 1992 Jun;6(3):237-43. doi: 10.1016/0890-8508(92)90022-p.
Synthetic oligonucleotides corresponding to a conserved 22 nucleotide sequence within the tandemly repeated mini-exon gene have been used to amplify a single gene-containing repeat from Trypanosoma cruzi and Trypanosoma rangeli, two morphologically similar organisms with overlapping hosts and geographical distribution but different pathogenicity in humans. The T. cruzi repeat is 582 nucleotides long and the T. rangeli repeat is 858 nucleotides. The two organisms may therefore be distinguished primarily by the electrophoretic mobilities of their respective amplification products. Confirmation of the diagnosis can be obtained by Southern blot analysis using species-specific DNA probes from the unique intergenic regions. We also present a diagnostic assay in which the unique intergenic regions are immobilized on nylon membranes and differentiation is based on hybridization with a digoxigenin-labelled PCR product.
与串联重复小外显子基因内一个保守的22个核苷酸序列相对应的合成寡核苷酸，已被用于从克氏锥虫和兰氏锥虫中扩增出一个含单个基因的重复序列。这两种锥虫在形态上相似，宿主和地理分布有重叠，但对人类的致病性不同。克氏锥虫的重复序列长582个核苷酸，兰氏锥虫的重复序列长858个核苷酸。因此，这两种生物主要可通过其各自扩增产物的电泳迁移率来区分。通过使用来自独特基因间隔区的种特异性DNA探针进行Southern印迹分析，可以确诊。我们还提出了一种诊断检测方法，其中独特的基因间隔区被固定在尼龙膜上，区分是基于与地高辛标记的PCR产物的杂交。
