Species-specific DNA probes for the identification of African trypanosomes in tsetse flies

Abstract

We have obtained 5 specific DNA probes for African trypanosomes of the subgenera Trypanozoon and Nannomonas. Each probe consists of one repeat unit of the major repetitive DNA (satellite DNA) of each species or intra-specific group. One probe hybridized with all members of subgenus Trypanozoon (except T. equiperdum which was not tested). In subgenus Nannomonas, one probe recognized T. simiae, but 3 probes were needed to identify all stocks of T. congolense available. Each of the 3 latter probes recognized trypanosomes from one of the 3 major groups of T. congolense previously defined by isoenzyme characterization, i.e. savannah, forest and Kenya coast types. As few as 100 trypanosomes could be unequivocally identified by dot blot hybridization and individual trypanosomes could be identified by in situ hybridization. We show how this simple methodology can be used in the field for the identification of immature and mature trypanosome infections in tsetse.

Trypanosoma congolense: the distribution of enzymic variants in East and West Africa

“…On the other hand, the cladogram, because it used weighted data, emphasizes the difference clearly. It also brings the 'Uganda savanna' group closer to the rest of the members of the East/West African savanna group, clustering both together into Cluster 1, thus supporting the DNA probe specificity (Gibson et al 1988). It is wise therefore to use two or more methods, in data analysis, and to evaluate the different groups against other evidence.…”

Kinetoplast DNA minicircles are inherited from both parents in genetic crosses of Trypanosoma brucei

“…Gels were blotted in the standard way (Southern 1975) and nitrocellulose ®lters were hybridised as previously described (Gibson et al 1988) with the oligo-labeled DNA probes A±E indicated in Fig. 1.…”

Analysis of isolates within species of anuran trypanosomes using random amplified polymorphic DNA

“…However, many species of Try-Z.-R. Lun 9 S.S. Desser (~) Department of Zoology, University of Toronto, Toronto, Ontario, Canada M5S 1A1 Fax (416) 978-8532 panosoma infecting anurans from different geographic areas are morphologically similar, and some species exhibit polymorphism in the vertebrate host, resulting in considerable taxonomic confusion (Martin 1993). Analysis of isoenzymes (Gibson et al 1980;Martin et al 1992a;Stevens et al 1992;Mathieu-Daude and Tibayrenc 1994), kinetoplast DNA (Borst et al 1981;Masiga and Gibson 1990), restriction-fragment-length polymorphism (RFLP; Paindavoine et al 1986;Gibson et al 1988;Hide et al 1990), molecular karyotypes (Giannini et al 1986), and random amplified polymorphic DNA (RAPD) have been used to investigate the genetic variation, molecular epidemiology, and identification of trypanosomes of mammals (Myler 1993), but there are few biochemical and molecular data on anuran trypanosomes.…”